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OpenMS
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Fragment-index-based peptide database search algorithm (experimental). More...
#include <OpenMS/ANALYSIS/ID/ProSEAlgorithm.h>
Classes | |
| struct | AnnotatedHit_ |
| Slimmer structure as storing all scored candidates in PeptideHit objects takes too much space. More... | |
| struct | CalibrationResult_ |
| Result of a calibration pass. More... | |
| struct | CandidatePoolStats_ |
| Running summary of the complete candidate pool of one spectrum. More... | |
| struct | CoIsolatedPSM_ |
| The PSM of a precursor co-isolated with the selected one (chimeric:enabled), until its residual spectrum is built. More... | |
| struct | DecoyStrategy_ |
| Resolved decoy handling for one concrete input database. More... | |
| struct | DeisotopingSettings_ |
| MS2 deisotoping rule of preprocessSpectra_(). More... | |
| struct | LocalFragmentEvidence_ |
| Local fragment evidence of one candidate (annotate:local_fragment_evidence). More... | |
| struct | MergedResult |
| The pooled evaluation of a multi-file search (the ProSE tool's -out_merged). More... | |
| struct | MultiFileSearchResult |
| Multi-file search result bundle. More... | |
| struct | PercolatorRescoring |
| Outcome of the Percolator rescoring of one search run (see rescoreWithPercolator() and the parameter 'rescore'). More... | |
| struct | PercolatorTraining |
| Options of Percolator's training for the rescoring (the parameters 'rescoring:...'; the defaults are ProSE's training: all PSMs, 10 iterations) More... | |
| struct | ProteinMapping_ |
| The protein occurrences of the sequences of a search's hits (peptide:protein_mapping = index). More... | |
| struct | ReuseRun_ |
| rescore_reuse: a run that trains models: its index in the search (the random halves depend on it) and its PSMs More... | |
| struct | RunStatistics |
| Per-run identification statistics for the end-of-search report. More... | |
| struct | SearchContext |
| Prepared per-database state shared across multiple spectrum files. More... | |
| struct | SearchResult |
| Comprehensive search result including modification analysis. More... | |
| struct | SharedSearchStats |
| Configuration, database and fragment-index facts shared across all input files of one ProSE invocation. More... | |
| struct | SpectraCounts_ |
Public Types | |
| enum class | ExitCodes { EXECUTION_OK , INPUT_FILE_EMPTY , UNEXPECTED_RESULT , UNKNOWN_ERROR , ILLEGAL_PARAMETERS } |
| Exit codes. More... | |
| using | FileResultConsumer = std::function< void(Size, const SearchResult &)> |
| The consumer of the files' results (setFileResultConsumer()): file index, result. Must only read the result. | |
 Public Types inherited from ProgressLogger | |
| enum | LogType { CMD , GUI , NONE } |
| Possible log types. More... | |
Public Member Functions | |
| ProSEAlgorithm () | |
| ExitCodes | search (const std::string &in_spectra, const std::string &in_db, std::vector< ProteinIdentification > &prot_ids, PeptideIdentificationList &pep_ids) const |
| Search spectra in a spectrum file (mzML or Bruker .d) against a protein database using an FI-backed workflow. | |
| void | annotatePsmQValues (PeptideIdentificationList &peptide_ids) const |
| Annotate PSM q-values by target-decoy competition of the best hit of each spectrum, as FDR:PSM does. | |
| bool | applyDeferredPsmFDR (const std::vector< ProteinIdentification > &protein_ids, PeptideIdentificationList &peptide_ids, bool rescored, double psm_fdr, PeptideIdentificationList *inference_only_psms=nullptr) const |
| Apply the PSM-level FDR to PSMs that were rescored separately, e.g. with rescoreWithPercolator() on the result of a search with 'rescore' and the FDR filters off. | |
| SearchResult | searchWithModificationAnalysis (const std::string &in_spectra, const std::string &in_db, const std::string &output_base_name="") const |
| Search with comprehensive results including modification analysis tables. | |
| ExitCodes | search (PeakMap &spectra, const std::vector< FASTAFile::FASTAEntry > &fasta_db, std::vector< ProteinIdentification > &prot_ids, PeptideIdentificationList &pep_ids) const |
| In-memory search: search spectra against a protein database without file I/O. | |
| SearchContext | prepareContext (const std::vector< FASTAFile::FASTAEntry > &fasta_db) const |
| Build a SearchContext (decoy-augmented database + FragmentIndex) for reuse. | |
| SearchContext | prepareContext (const std::vector< FASTAFile::FASTAEntry > &fasta_db, bool electron_ions) const |
| Build a SearchContext whose FragmentIndex also holds c and z+1 ions. | |
| ExitCodes | search (PeakMap &spectra, SearchContext &ctx, std::vector< ProteinIdentification > &prot_ids, PeptideIdentificationList &pep_ids) const |
| In-memory search using a pre-built SearchContext. | |
| SearchResult | searchWithModificationAnalysis (PeakMap &spectra, const std::vector< FASTAFile::FASTAEntry > &fasta_db, const std::string &output_base_name="") const |
| In-memory search with modification analysis: no file I/O required. | |
| MultiFileSearchResult | searchWithModificationAnalysis (const std::vector< std::string > &in_spectra_files, const std::vector< FASTAFile::FASTAEntry > &fasta_db, const std::vector< std::string > &output_base_names={}, const std::string &aggregate_base_name="", bool build_pooled_aggregate=true) const |
| Multi-file search with modification analysis (in-memory FASTA). | |
| MultiFileSearchResult | searchWithModificationAnalysis (const std::vector< std::string > &in_spectra_files, const std::string &in_db, const std::vector< std::string > &output_base_names={}, const std::string &aggregate_base_name="", bool build_pooled_aggregate=true) const |
| Multi-file search with modification analysis (FASTA file path). | |
| MergedResult | mergeFileResults (MultiFileSearchResult &result, const StringList &in_spectra_files, bool remove_decoys=true) const |
| Pools result (moving its PSMs out), infers proteins across files, applies FDR:protein of this instance with picked-protein FDR if decoy proteins were identified and then, by default, removes the decoys. | |
| void | setParameterNamePrefix (const std::string &prefix) |
| void | setKeepPreFdrPsms (bool keep) |
| void | setFileResultConsumer (FileResultConsumer consumer) |
Lets the multi-file searchWithModificationAnalysis() overloads hand each file's result to consumer as soon as it is final, e.g. to write it while the next file is searched. | |
| Public Member Functions inherited from DefaultParamHandler | |
| DefaultParamHandler (const std::string &name) | |
| Constructor with name that is displayed in error messages. | |
| DefaultParamHandler (const DefaultParamHandler &rhs) | |
| Copy constructor. | |
| virtual | ~DefaultParamHandler () |
| Destructor. | |
| DefaultParamHandler & | operator= (const DefaultParamHandler &rhs) |
| Assignment operator. | |
| virtual bool | operator== (const DefaultParamHandler &rhs) const |
| Equality operator. | |
| void | setParameters (const Param ¶m) |
| Sets the parameters. | |
| const Param & | getParameters () const |
| Non-mutable access to the parameters. | |
| const Param & | getDefaults () const |
| Non-mutable access to the default parameters. | |
| const std::string & | getName () const |
| Non-mutable access to the name. | |
| void | setName (const std::string &name) |
| Mutable access to the name. | |
| const std::vector< std::string > & | getSubsections () const |
| Non-mutable access to the registered subsections. | |
| Public Member Functions inherited from ProgressLogger | |
| ProgressLogger () | |
| Constructor. | |
| virtual | ~ProgressLogger () |
| Destructor. | |
| ProgressLogger (const ProgressLogger &other) | |
| Copy constructor. | |
| ProgressLogger & | operator= (const ProgressLogger &other) |
| Assignment Operator. | |
| void | setLogType (LogType type) const |
| Sets the progress log that should be used. The default type is NONE! | |
| LogType | getLogType () const |
| Returns the type of progress log being used. | |
| void | setLogger (ProgressLoggerImpl *logger) |
| Sets the logger to be used for progress logging. | |
| void | startProgress (SignedSize begin, SignedSize end, const std::string &label) const |
| Initializes the progress display. | |
| void | setProgress (SignedSize value) const |
| Sets the current progress. | |
| void | endProgress (UInt64 bytes_processed=0) const |
| void | nextProgress () const |
| increment progress by 1 (according to range begin-end) | |
Static Public Member Functions | |
| static void | stratumQValues (PeptideIdentificationList &peptide_ids) |
| rescoring:stratum_q_values: grouped target-decoy competition on the best hit of every spectrum that has the meta value percolator_score; stratum 'protein_nterm' (an evidence at a protein N-terminus, or only Met-clipped evidences: aa_before M at start 1) or 'other'; q = (decoys + 1) / max(targets, 1) in percolator_score order (ties: decoys first), monotone. Sets the meta values 'stratum' and 'stratum_q_value'; a best hit without percolator_score gets only 'stratum_q_value' = 1. | |
| static bool | reuseTrainingDraw (UInt64 seed, Size file, Size spectrum, double fraction) |
rescore_reuse: whether spectrum spectrum (its index in the run) of file file is drawn for the training of the models at the sampling fraction fraction (rescore_reuse:max_train) | |
| static bool | reuseCalibrationDraw (UInt64 seed, Size file, Size spectrum, double fraction) |
| static bool | applyCompleteSetProteinFDR (std::vector< ProteinIdentification > &protein_ids, PeptideIdentificationList &peptide_ids, const std::string &decoy_string, bool decoy_is_prefix, double protein_fdr, bool remove_decoys, const PeptideIdentificationList &inference_only_psms) |
| Finalize protein-level FDR on a COMPLETE protein set (a single input file, or a merged cross-file aggregate). | |
| static std::string | inferProteins (std::vector< ProteinIdentification > &protein_ids, const PeptideIdentificationList &peptide_ids, const PeptideIdentificationList &inference_only_psms=PeptideIdentificationList()) |
| The protein inference of the protein FDR (applyCompleteSetProteinFDR()) and of mergeFileResults(). | |
| static void | removeUnreportedProteins (std::vector< ProteinIdentification > &protein_ids, const PeptideIdentificationList &peptide_ids, const PeptideIdentificationList &also_referencing=PeptideIdentificationList()) |
| Removes the proteins that no PSM references, and them from the protein groups. | |
| static Size | removeDuplicateProteins (std::vector< FASTAFile::FASTAEntry > &db, const std::string &source) |
| Removes the repeated entries of a protein database, which searching would report twice (e.g. a FASTA file concatenated twice, or contaminants added twice). | |
| static PercolatorRescoring | rescoreWithPercolator (std::vector< ProteinIdentification > &protein_ids, PeptideIdentificationList &peptide_ids, int threads, Size max_train_psms=0) |
| Rescore the PSMs of one search run in-process with Percolator, as the parameter 'rescore' does in every search. | |
| static std::string | pinEnzymeName (const std::string &enzyme) |
The name of enzyme as PercolatorInfile computes the enzN/enzC/enzInt features of a .pin. | |
| static std::string | pinFeatureEnzyme (const std::string &enzyme, bool fixed_enzyme_termini) |
The enzyme argument of PercolatorInfile (store(), the in-process rescoring) for the enzN/enzC/enzInt features of the search's enzyme: its Percolator name (pinEnzymeName(); a ProteaseDB enzyme without one keeps its name, so that PercolatorInfile uses its cleavage rule) with "+termini" (fixed_enzyme_termini: protein termini and the position after an initial Met are cleavage sites) or "+keep_termini" (the features of ProSE int15, the default). | |
| static PeakMap | loadSpectra (const std::string &filename, const std::function< bool(const MSSpectrum &)> &keep={}) |
| The MS2 spectra of a spectrum file as the searches of this class read them. | |
| static void | splitMergedPsms (MergedResult &merged, MultiFileSearchResult &result) |
| Makes the PSMs of each file of a multi-file search its part of the pooled result (mergeFileResults()), so that per-file outputs agree with the pooled one. | |
| static void | updateFinalStats (RunStatistics &stats, const PeptideIdentificationList &peptide_ids, const std::string &enzyme, bool fdr_applied) |
| static void | releaseInParallel (SearchResult &result) |
| static std::string | renderTotalTime (const SharedSearchStats &sh) |
The total time of a run, by part: "total <s>s | search of the file(s) <s>s | outputs <s>s
(of which -qpx_extended <s>s)", from sh's seconds_total, seconds_output and seconds_qpx_extended (the last part only if positive). The end of the report of renderRunSummary(), and of ProSE's report of several files. | |
| static void | renderRunSummary (const RunStatistics &stats, const SharedSearchStats &shared, const OpenSearchModificationAnalysis::OpenSearchAnalysisResult &mod_analysis, bool is_open_search, std::ostream &os) |
| static void | renderModificationSummary (const OpenSearchModificationAnalysis::OpenSearchAnalysisResult &mod_analysis, std::ostream &os) |
| static std::string | renderRunSummaryYaml (const MultiFileSearchResult &mfres, const std::vector< std::pair< std::string, std::vector< std::string > > > &manifest, Size files_failed, Size files_total) |
| Static Public Member Functions inherited from DefaultParamHandler | |
| static void | writeParametersToMetaValues (const Param &write_this, MetaInfoInterface &write_here, const std::string &key_prefix="") |
| Writes all parameters to meta values. | |
Static Public Attributes | |
| static constexpr const char * | CHIMERIC_RANK = "chimeric_rank" |
| static constexpr double | MIN_DECOYS_PER_TARGET = 0.5 |
| static constexpr double | MAX_DECOYS_PER_TARGET = 2.0 |
| See MIN_DECOYS_PER_TARGET. | |
Protected Types | |
| enum class | DecoyMode_ { AUTO , GENERATE , IGNORE } |
| How decoys are obtained/recognised for a search (parameter "decoys"). More... | |
Protected Member Functions | |
| void | updateMembers_ () override |
| This method is used to update extra member variables at the end of the setParameters() method. | |
| void | annotateIonPriors_ (const PeakMap &spectra, const FragmentIonLikelihoodModel::PeakLists &evidence, std::vector< ProteinIdentification > &protein_ids, PeptideIdentificationList &peptide_ids) const |
| Learn per-run fragment ion likelihoods from confident PSMs and annotate every hit with them. | |
| DecoyStrategy_ | resolveDecoyStrategy_ (const std::vector< FASTAFile::FASTAEntry > &db) const |
Decide how to obtain/recognise decoys for db. | |
| Param | fragmentIndexParameters_ (bool electron_ions=false) const |
| ProSE parameters made safe to hand to a FragmentIndex. | |
| SpectrumGenerators_ | spectrumGenerators_ () const |
| Generators for scoring, annotation and calibration, configured from the ions:* parameters. | |
| Size | countElectronActivated_ (const PeakMap &spectra) const |
| Number of spectra that ions:by_activation also scores with c and z+1 ions (0 if it is off) | |
| std::vector< FASTAFile::FASTAEntry > | buildDecoyAugmentedDB_ (const std::vector< FASTAFile::FASTAEntry > &fasta_db, const DecoyStrategy_ &strategy) const |
Build the searched database according to strategy. | |
| std::vector< FASTAFile::FASTAEntry > | buildDecoyAugmentedDB_ (std::vector< FASTAFile::FASTAEntry > &&fasta_db, const DecoyStrategy_ &strategy) const |
As above, but takes the entries of fasta_db over instead of copying them. | |
| MultiFileSearchResult | searchWithModificationAnalysis_ (const std::vector< std::string > &in_spectra_files, const std::vector< FASTAFile::FASTAEntry > &fasta_db_given, const std::string *in_db, const std::vector< std::string > &output_base_names, const std::string &aggregate_base_name, bool build_pooled_aggregate) const |
| void | reportDecoyParity_ (const SearchContext &ctx) const |
| SearchContext | prepareContext_ (std::vector< FASTAFile::FASTAEntry > &&fasta_db, bool electron_ions, const std::function< const PeakMap *(Size)> &searched_spectra={}) const |
| bool | restrictIndexToSpectra_ () const |
| void | buildFragmentIndex_ (SearchContext &ctx, const Param ¶ms, const std::function< const PeakMap *(Size)> &searched_spectra) const |
| bool | chimericSearch_ () const |
| void | searchCoIsolated_ (const PeakMap &spectra, const std::vector< std::vector< AnnotatedHit_ > > &annotated_hits, FragmentIndex &fi, const std::vector< FASTAFile::FASTAEntry > &db, const SpectrumGenerators_ &generators, double fragment_tol, bool fragment_tol_ppm, bool open_search_mode, std::vector< CoIsolatedPSM_ > &psms, std::vector< std::vector< AnnotatedHit_ > > &hits, std::vector< CandidatePoolStats_ > &pool_stats) const |
| Search the precursors co-isolated with the selected one in what the spectrum's other PSMs leave of it (chimeric:enabled), after the andes chimeric cascade. | |
| std::vector< FASTAFile::FASTAEntry > | buildCalibrationSample_ (const std::vector< FASTAFile::FASTAEntry > &full_db) const |
| Build a strided protein sample for chunked calibration. | |
| ExitCodes | searchChunked_ (PeakMap &spectra, std::vector< FASTAFile::FASTAEntry > &full_db, const DecoyStrategy_ &strategy, std::vector< ProteinIdentification > &protein_ids, PeptideIdentificationList &peptide_ids) const |
| Chunked database search implementation. | |
| void | scoreSpectraAgainstIndex_ (const PeakMap &spectra, FragmentIndex &fi, const std::vector< FASTAFile::FASTAEntry > &db, const SpectrumGenerators_ &generators, double effective_fragment_tol, bool fragment_mass_tolerance_unit_ppm, bool open_search_mode, std::vector< std::vector< AnnotatedHit_ > > &annotated_hits, std::vector< CandidatePoolStats_ > &pool_stats, const std::string &progress_label, const PeakMap *query_spectra=nullptr) const |
| Score all spectra against one FragmentIndex. | |
| void | postProcessHits_ (const PeakMap &exp, std::vector< std::vector< ProSEAlgorithm::AnnotatedHit_ > > &annotated_hits, const std::vector< CandidatePoolStats_ > &pool_stats, std::vector< ProteinIdentification > &protein_ids, PeptideIdentificationList &peptide_ids, Size top_hits, const StringList &modifications_fixed, const StringList &modifications_variable, Int peptide_missed_cleavages, double precursor_mass_tolerance, double fragment_mass_tolerance, const std::string &precursor_mass_tolerance_unit_ppm, const std::string &fragment_mass_tolerance_unit_ppm, const Int precursor_min_charge, const Int precursor_max_charge, const std::string &enzyme, const std::string &database_name, const PeakMap *evidence_spectra=nullptr, const std::vector< Size > *evidence_parent=nullptr, bool defer_peak_annotations=false) const |
| Filter and annotate search results. | |
| void | annotateFragmentPeaks_ (const PeakMap &exp, PeptideIdentificationList &peptide_ids, double fragment_mass_tolerance, const std::string &fragment_mass_tolerance_unit_ppm) const |
The fragment annotations that postProcessHits_() leaves out with defer_peak_annotations, for the hits of peptide_ids (by their "scan_index" in exp), computed as postProcessHits_() computes them. | |
| void | annotatePeptDeepFeatures_ (const PeakMap &spectra, std::vector< ProteinIdentification > &protein_ids, PeptideIdentificationList &peptide_ids) const |
| Adds PeptDeep-prediction-derived rescoring features to the PSMs. | |
| int | scoringMaxCharge_ (int precursor_charge) const |
| Fragment charges that score a candidate of the given precursor charge; also used by the calibration pass. | |
| Size | rescoreTopHits_ () const |
| The hits per spectrum that Percolator rescores: rescoring:top_hits, or report:top_hits if that is 0. | |
| Size | searchTopHits_ () const |
| std::string | paramName_ (const std::string &name) const |
Helper: name as the user sets the parameter (setParameterNamePrefix()), for messages. | |
| bool | keepPreRescoringPsms_ () const |
| Whether a search keeps the PSMs of each file before the rescoring (SearchResult::pre_fdr_peptide_ids) | |
| Size | transferQuota_ () const |
| std::shared_ptr< const ReuseModel_ > | trainReuseModel_ (const std::vector< ReuseRun_ > &runs, int threads) const |
| PercolatorRescoring | rescoreWithReuseModel_ (std::vector< ProteinIdentification > &protein_ids, PeptideIdentificationList &peptide_ids, Size file_index, const ReuseModel_ &model, bool check_shift, int threads, std::vector< std::vector< PeptideHit > > *dropped) const |
| void | applyRescoredPsmFDR_ (std::vector< ProteinIdentification > &protein_ids, PeptideIdentificationList &peptide_ids, bool has_decoys, RunStatistics &stats, bool rescored, PeptideIdentificationList *inference_only_psms) const |
| void | finishDeferredRescoring_ (MultiFileSearchResult &mfres, const std::vector< Size > &deferred) const |
| double | computeModMatchTolerance_ () const |
| CalibrationResult_ | runCalibrationPass_ (PeakMap &spectra, FragmentIndex &fragment_index, const std::vector< FASTAFile::FASTAEntry > &db, const PeakMap *query_spectra=nullptr) const |
| Run a fast calibration pass on a subset of spectra to estimate mass accuracy. | |
| PercolatorRescoring | applyPsmFDR_ (std::vector< ProteinIdentification > &protein_ids, PeptideIdentificationList &peptide_ids, bool has_decoys, RunStatistics &stats, PeptideIdentificationList *pre_fdr_psms=nullptr, PeptideIdentificationList *inference_only_psms=nullptr) const |
| void | rescoreFilesTogether_ (MultiFileSearchResult &mfres, const std::vector< std::string > &in_spectra_files) const |
| The end of a multi-file search with 'rescore' (with pool_rescoring_), for the files with PSMs that Percolator could not rescore on their own (e.g. fewer than 100 PSMs, or no decoy PSM). | |
| void | collectInferenceOnlyPsms_ (const PeptideIdentificationList &peptide_ids, double psm_fdr, PeptideIdentificationList &inference_only_psms) const |
| void | collectRunStatistics_ (const PeakMap &spectra, const std::vector< ProteinIdentification > &protein_ids, const PeptideIdentificationList &peptide_ids, RunStatistics &stats) const |
| SpectraCounts_ | countSpectra_ (const PeakMap &spectra) const |
Helper: the SpectraCounts_ of spectra. | |
| void | preprocessSearchedSpectra_ (PeakMap &spectra, const std::string &input, FragmentIonLikelihoodModel::PeakLists &ion_evidence, PeakMap *evidence_spectra, PeakMap *query_spectra) const |
| Protected Member Functions inherited from DefaultParamHandler | |
| void | defaultsToParam_ () |
| Updates the parameters after the defaults have been set in the constructor. | |
Static Protected Member Functions | |
| static void | preprocessSpectra_ (PeakMap &exp, double fragment_mass_tolerance, bool fragment_mass_tolerance_unit_ppm, bool deisotope_requested, Size peaks_keep_n, Int peaks_window_top, const std::string &window_type, const DeisotopingSettings_ &deisotoping, FragmentIonLikelihoodModel::PeakLists *ion_evidence=nullptr, bool ion_evidence_scored_peaks=false, PeakMap *evidence_spectra=nullptr, PeakMap *query_spectra=nullptr) |
| filter, deisotope, decharge spectra | |
| static void | preprocessSpectra_ (PeakMap &exp, double fragment_mass_tolerance, bool fragment_mass_tolerance_unit_ppm, bool deisotope_requested, Size peaks_keep_n, Int peaks_window_top, const std::string &window_type="auto") |
| preprocessSpectra_() with the DeisotopingSettings_ defaults (the behaviour before its parameters existed) | |
| static AASequence | reversedNoiseSequence_ (const AASequence &sequence) |
| Reversed sequence keeping the C-terminal residue and every residue's modification: the noise hypothesis of a confident PSM. | |
| static void | filterLocalPeaks_ (MSSpectrum &spectrum, Size peaks_per_window) |
| Keep the strongest peaks in each non-overlapping 100 Da window, including a short final window. | |
| static std::vector< double > | localPeakDensities_ (const MSSpectrum &spectrum) |
| Peak counts per Da in a fixed +/-50 m/z window around each peak of a sorted spectrum. | |
| static LocalFragmentEvidence_ | localFragmentEvidence_ (const MSSpectrum &spectrum, const MSSpectrum &theoretical, const std::vector< double > &densities, double tolerance, bool ppm) |
| Compute local fragment-match evidence on the spectrum before window/top-N peak filtering. | |
| static LocalFragmentEvidence_ | localFragmentEvidence_ (const MSSpectrum &spectrum, const MSSpectrum &theoretical, int max_charge, const std::vector< double > &densities, double tolerance, bool ppm, std::vector< double > &alternatives) |
| static LocalFragmentEvidence_ | localFragmentEvidence_ (const MSSpectrum &spectrum, const MSSpectrum &theoretical, const std::vector< Int > &charges, int max_charge, const std::vector< double > &densities, double tolerance, bool ppm, std::vector< double > &alternatives) |
localFragmentEvidence_() with the charges of the peaks of theoretical given (charges, one per peak) | |
| static bool | isElectronActivated_ (const MSSpectrum &spectrum) |
True if the precursor of spectrum was activated by electrons (ETD, ECD, EThcD or ETciD) | |
| static double | calibrationPassWindow_ (double configured, bool ppm) |
The window the calibration pass searches with, from the configured one (ppm or Da) | |
| static MSSpectrum | residualSpectrum_ (const MSSpectrum &spectrum, const std::vector< UInt32 > &removed, const Precursor &precursor) |
| static std::vector< SignedSize > | scoringOrder_ (const PeakMap &spectra, const PeakMap *query_spectra) |
The order in which scoreSpectraAgainstIndex_() scores the spectra: the scan indices sorted by the neutral precursor mass of the first precursor (of query_spectra if given; an unknown charge counts as 1), ties by scan index; spectra without a finite mass at the end in scan order. | |
| static std::vector< Size > | transferSample_ (const PeptideIdentificationList &ids, Size max_psms, Size hits_per_spectrum) |
The spectra (positions in ids) of a seeded random sample of whole spectra with at most max_psms PSMs, counting at most hits_per_spectrum hits of a spectrum; in ascending order. A spectrum's place in the draw depends only on the spectrum (its reference, retention time and precursor m/z), so the sample of a list is also the sample of any of its sublists that contain it (the copy of a file reduced to it after the rescoring, see applyPsmFDR_()), and it is the same on every platform. Spectra without hits are left out. | |
| static PercolatorRescoring | rescoreWithPercolator_ (std::vector< ProteinIdentification > &protein_ids, PeptideIdentificationList &peptide_ids, int threads, std::vector< std::vector< PeptideHit > > *dropped, const PercolatorTraining &training, const Param &extra, const ReuseScoring_ *reuse=nullptr) |
| static ProteinMapping_ | buildProteinMapping_ (const FragmentIndex &index, const std::vector< FASTAFile::FASTAEntry > &db, std::vector< UInt32 > candidates, const Param &indexer_parameters) |
Collects the protein occurrences of the sequences of the index peptides candidates. | |
| static bool | applyProteinMapping_ (const ProteinMapping_ &mapping, const std::vector< FASTAFile::FASTAEntry > &db, const Param &indexer_parameters, std::vector< ProteinIdentification > &protein_ids, PeptideIdentificationList &peptide_ids, std::string &fallback_reason) |
Writes what PeptideIndexing::run(db, protein_ids, peptide_ids) with indexer_parameters writes: the evidences, target_decoy and protein_references of every hit, the protein hits and the PeptideIndexer:* search parameters. | |
| static bool | accessionHasDecoyMarker_ (const std::string &accession, const std::string &marker, bool is_prefix) |
| static void | removeProteinsReferencedByNeither_ (std::vector< ProteinIdentification > &protein_ids, const PeptideIdentificationList &peptide_ids, const PeptideIdentificationList &also_referencing) |
| static bool | useSearchEngineScore_ (PeptideIdentificationList &ids) |
| static Size | countDecoyEntries_ (const std::vector< FASTAFile::FASTAEntry > &db, const std::string &marker, bool is_prefix) |
Helper: the entries of db whose accession carries the decoy marker (accessionHasDecoyMarker_()) | |
| static void | poolSearchedFiles_ (std::vector< SearchResult > &per_file, bool move_psms, const StringList &in_spectra_files, std::vector< ProteinIdentification > &protein_ids, PeptideIdentificationList &peptide_ids) |
| static bool | searched_ (const SearchResult &pf) |
Helper: whether the file of pf was searched (exit code EXECUTION_OK, with a run) | |
| static int | searchedFileOf_ (const PeptideIdentification &id, const std::vector< SearchResult > &per_file) |
| static void | returnPooledPsms_ (PeptideIdentificationList &pooled, std::vector< SearchResult > &per_file) |
| static void | capturePreFdrStats_ (const PeptideIdentificationList &peptide_ids, RunStatistics &stats) |
| static double | maxRetainedScore_ (const PeptideIdentificationList &peptide_ids) |
| static void | keepTopHits_ (PeptideIdentificationList &peptide_ids, Size n) |
| static void | markRescoredByPercolator_ (std::vector< ProteinIdentification > &protein_ids, const Param &pp, const PercolatorTraining &training, const Percolator *perc=nullptr, const ReuseScoring_ *reuse=nullptr) |
| static bool | hasPSMs_ (const PeptideIdentificationList &ids) |
Helper: whether a PSM of ids has a hit. | |
| static void | tdcQValues_ (PeptideIdentificationList &peptide_ids, bool plus_one=false) |
Protected Attributes | |
| bool | peptdeep_enable_ {false} |
| std::string | peptdeep_ms2_model_ |
| std::string | peptdeep_rt_model_ |
| std::string | peptdeep_instrument_ {"QE"} |
| double | peptdeep_nce_ {-1.0} |
| std::string | peptdeep_rt_model_type_ {"b_spline"} |
| int | peptdeep_batch_size_ {500} |
| double | precursor_mass_tolerance_lower_ {10.0} |
| positive magnitude (default matches the param) | |
| double | precursor_mass_tolerance_upper_ {10.0} |
| positive magnitude (default matches the param) | |
| std::string | precursor_mass_tolerance_unit_ {"ppm"} |
| Size | precursor_min_charge_ |
| Size | precursor_max_charge_ |
| double | fragment_mass_tolerance_ |
| std::string | fragment_mass_tolerance_unit_ |
| bool | deisotope_requested_ {true} |
| bool | scoring_multiple_charges_ {false} |
| Resolved scoring:fragment_charges: score charges above 1. | |
| int | scoring_max_charge_ {2} |
| Upper fragment charge bound (fragment:max_charge) | |
| bool | mass_accuracy_score_ {false} |
| Resolved scoring:method: mass-accuracy weighted HyperScore. | |
| double | mass_error_sd_ppm_ {7.0} |
| Width of the mass-accuracy kernel in ppm (scoring:mass_error_sd) | |
| Size | peaks_keep_n_ {0} |
| NLargest cap on MS2 peaks before scoring; 0 = resolution-aware auto (peaks:keep_n) | |
| Int | peaks_window_top_ {20} |
| WindowMower peaks-per-100Da before scoring (peaks:window_top) | |
| std::string | peaks_window_type_ {"auto"} |
| Resolution-aware treatment of the final peak window. | |
| DeisotopingSettings_ | deisotoping_ |
| MS2 deisotoping rule (fragment:deisotope_*) | |
| bool | query_raw_spectrum_ {false} |
| Retrieve candidates with the spectrum before deisotoping and local filtering (fragment:query_spectrum) | |
| StringList | modifications_fixed_ |
| StringList | modifications_variable_ |
| Size | modifications_max_variable_mods_per_peptide_ |
| std::string | enzyme_ |
| DecoyMode_ | decoy_mode_ {DecoyMode_::AUTO} |
| std::string | decoy_prefix_ |
| bool | decoy_keep_termini_ = false |
| decoy_method 'keep_termini' | |
| bool | decoy_parity_report_ = false |
| decoy_parity_report | |
| bool | terminal_features_ = false |
| rescoring:terminal_features (effective) | |
| bool | stratum_q_values_ = false |
| rescoring:stratum_q_values (effective) | |
| bool | nterm_options_disabled_ = false |
| both switched off by buildDecoyAugmentedDB_() | |
| bool | rescore_ {true} |
| rescore: Percolator rescoring of every run before its PSM-level FDR | |
| PercolatorTraining | percolator_training_ |
| rescoring:...: options of Percolator's training | |
| Param | percolator_extra_ |
| rescore_method 'reset': Percolator's RESET parameters (empty: Percolator) | |
| std::string | reset_run_name_ |
| spectra file of the run being rescored (names its RESET stem table) | |
| std::string | reuse_mode_ {"per_file"} |
| rescore_reuse:mode | |
| Size | reuse_max_train_ {250000} |
| rescore_reuse:max_train | |
| Size | reuse_static_files_ {1} |
| rescore_reuse:static_files | |
| double | reuse_max_shift_ {0.5} |
| rescore_reuse:max_shift | |
| Size | max_train_psms_ {500000} |
| percolator:max_train_psms (0: no cap) | |
| double | fdr_psm_ {0.0} |
| bool | fdr_psm_by_scored_charges_ {true} |
| FDR:PSM_groups: separate competitions per number of scored fragment charges. | |
| double | fdr_protein_ {0.0} |
| double | fdr_protein_inference_psm_ {0.1} |
| FDR:protein_inference_PSM: q-value cutoff of the PSMs of the protein inference. | |
| StringList | annotate_psm_ |
| bool | self_trained_ion_priors_ {false} |
| annotate:self_trained_ion_priors | |
| FragmentIonLikelihoodModel::ContextSet | ion_prior_model_ {FragmentIonLikelihoodModel::ContextSet::RICH} |
| annotate:ion_prior_model | |
| bool | ion_prior_scored_peaks_ {false} |
| annotate:ion_prior_peaks == scored | |
| int | ion_prior_max_fragment_charge_ {2} |
| annotate:ion_prior_max_fragment_charge (capped at precursor charge - 1) | |
| double | ion_prior_train_fdr_ {0.01} |
| annotate:ion_prior_train_fdr: TDC q-value threshold of the training PSMs | |
| Size | ion_prior_min_psms_ {100} |
| annotate:ion_prior_min_psms: training PSMs required per fold | |
| bool | ion_prior_feature_llr_ {true} |
| annotate:ion_prior_features holds ion_prior_llr | |
| bool | ion_prior_feature_explained_ {true} |
| annotate:ion_prior_features holds ion_prior_explained | |
| bool | ion_prior_feature_topk_ {false} |
| annotate:ion_prior_features holds ion_prior_topk_observed | |
| Size | peptide_min_size_ |
| Size | peptide_max_size_ |
| Size | peptide_missed_cleavages_ |
| EnzymaticDigestion::Specificity | peptide_enzyme_specificity_ {EnzymaticDigestion::SPEC_FULL} |
| Size | report_top_hits_ |
| Size | rescore_top_hits_ {0} |
| rescoring:top_hits: the hits per spectrum that the Percolator rescoring gets (0 = report_top_hits_) | |
| bool | isotope_error_observed_minus_theoretical_ {true} |
| bool | add_a_ions_ {false} |
| bool | add_b_ions_ {true} |
| bool | add_c_ions_ {false} |
| bool | add_x_ions_ {false} |
| bool | add_y_ions_ {true} |
| bool | add_z_ions_ {false} |
| bool | add_zp1_ions_ {false} |
| bool | ions_by_activation_ {true} |
| add c and z+1 ions for electron-activated spectra | |
| bool | protein_mapping_from_index_ {true} |
| peptide:protein_mapping = index | |
| Size | database_chunk_size_ {0} |
| 0 = disabled; >0 = chunk DB into groups of this many proteins | |
| bool | calibration_enabled_ {false} |
| double | calibration_subset_ratio_ {0.1} |
| bool | chimeric_enabled_ {false} |
| chimeric:enabled: also search the precursors co-isolated with the selected one | |
| Size | calibration_min_psms_ {50} |
| double | calibration_sigma_factor_ {4.0} |
| calibration:sigma_factor (fragment) | |
| double | calibration_precursor_sigma_factor_ {0.0} |
| calibration:precursor_sigma_factor (0: the precursor window is kept) | |
| double | calibration_min_precursor_tol_ {0.0} |
| calibration:min_precursor_tolerance (0: by unit) | |
| double | calibration_min_fragment_tol_ {0.0} |
| calibration:min_fragment_tolerance (0: by unit) | |
| CalibrationResult_ | last_calibration_result_ |
| RunStatistics | last_run_stats_ |
| PercolatorRescoring | last_percolator_ |
| bool | keep_pre_fdr_psms_ = false |
| see setKeepPreFdrPsms() | |
| FileResultConsumer | file_result_consumer_ |
| see setFileResultConsumer() | |
| std::string | parameter_name_prefix_ |
| bool | pool_rescoring_ = false |
| Size | pool_files_ = 0 |
| PeptideIdentificationList | pre_fdr_peptide_ids_ |
| std::shared_ptr< void > | released_hits_ |
| std::shared_ptr< const ReuseModel_ > | reuse_model_ |
| rescore_reuse: the models of a multi-file search ('static': trained on its first files; null otherwise) | |
| bool | reuse_defer_ = false |
| rescore_reuse: the current search defers its rescoring and FDR:PSM to finishDeferredRescoring_() | |
| Size | reuse_file_index_ = 0 |
| rescore_reuse: the index of the file of the current search in a multi-file search (0 otherwise) | |
| PeptideIdentificationList | inference_only_peptide_ids_ |
| double | last_mod_match_tolerance_used_ {-1.0} |
| Protected Attributes inherited from DefaultParamHandler | |
| Param | param_ |
| Container for current parameters. | |
| Param | defaults_ |
| Container for default parameters. This member should be filled in the constructor of derived classes! | |
| std::vector< std::string > | subsections_ |
| Container for registered subsections. This member should be filled in the constructor of derived classes! | |
| std::string | error_name_ |
| Name that is displayed in error messages during the parameter checking. | |
| bool | check_defaults_ |
| If this member is set to false no checking if parameters in done;. | |
| bool | warn_empty_defaults_ |
| If this member is set to false no warning is emitted when defaults are empty;. | |
| Protected Attributes inherited from ProgressLogger | |
| LogType | type_ |
| time_t | last_invoke_ |
| ProgressLoggerImpl * | current_logger_ |
Static Protected Attributes | |
| static constexpr const char * | SEARCH_ENGINE_SCORE = "ln(hyperscore)" |
| Static Protected Attributes inherited from ProgressLogger | |
| static std::atomic< int > | recursion_depth_ |
Private Member Functions | |
| bool | isOpenSearchMode_ () const |
| Helper function to determine if open search should be used based on tolerance. | |
Fragment-index-based peptide database search algorithm (experimental).
Provides a self-contained search engine that matches MS/MS spectra against a protein database using an FI (Fragment Index). Typical usage:
Notes:
| struct OpenMS::ProSEAlgorithm::CalibrationResult_ |
Result of a calibration pass.
Holds the estimated precursor and fragment tolerances computed from confident PSMs during the calibration pass. When success is false, the tolerance values are undefined and should not be used.
| Class Members | ||
|---|---|---|
| double | cal_lower {0} | calibrated lower magnitude (valid iff !extreme_bias && success) |
| double | cal_upper {0} | calibrated upper magnitude (valid iff !extreme_bias && success) |
| bool | extreme_bias {false} | zero spread of the precursor errors: the precursor window is kept (test observability) |
| double | fragment_coverage {0} | share of the matched fragment peaks inside the fragment tolerance |
| Size | fragment_peaks {0} | matched fragment peaks the fragment window was estimated from |
| double | fragment_shift {0} | median of the signed errors of the matched fragment peaks (covered by the window, not corrected) |
| double | fragment_sigma {0} | their robust standard deviation, 1.4826 * MAD |
| double | fragment_tolerance {0} | estimated fragment tolerance (same unit as configured) |
| double | precursor_coverage {0} | share of the calibration PSMs inside the precursor window median +/- k sigma |
| double | precursor_shift {0} | median of the signed precursor errors of the calibration PSMs (the mass offset) |
| double | precursor_sigma {0} | their robust standard deviation, 1.4826 * MAD |
| double | precursor_spread {0} | the larger side of the window before the caps, max(cal_lower, cal_upper) (diagnostic) |
| Size | psms {0} | calibration PSMs the windows were estimated from |
| bool | success {false} | true if enough PSMs were found for reliable estimation |
| struct OpenMS::ProSEAlgorithm::CoIsolatedPSM_ |
The PSM of a precursor co-isolated with the selected one (chimeric:enabled), until its residual spectrum is built.
| Class Members | ||
|---|---|---|
| Size | parent = 0 | index of its spectrum in the searched spectra |
| Precursor | precursor | the co-isolated precursor: that of the spectrum with its m/z and charge |
| vector< UInt32 > | removed | the peaks of the spectrum that its residual spectrum leaves out (ascending) |
| struct OpenMS::ProSEAlgorithm::DecoyStrategy_ |
Resolved decoy handling for one concrete input database.
Produced by resolveDecoyStrategy_() and consumed by buildDecoyAugmentedDB_() and the downstream PeptideIndexing / FDR steps, so the same decoys that are searched are also the ones scored.
| Class Members | ||
|---|---|---|
| string | decoy_string | effective marker for PeptideIndexing + protein FDR |
| bool | generate {false} | reverse target proteins to synthesise decoys |
| bool | have_decoys {false} | searched DB will contain decoys (FDR possible) |
| bool | is_prefix {true} | position of decoy_string |
| bool | strip_existing {false} | drop pre-existing decoy entries before searching |
| bool | strip_is_prefix {true} | position of strip_string |
| string | strip_string | marker of pre-existing decoys to strip |
| struct OpenMS::ProSEAlgorithm::DeisotopingSettings_ |
MS2 deisotoping rule of preprocessSpectra_().
Filled from the parameters fragment:deisotope_* by updateMembers_(), whose defaults are the Sage-like rule (min_peaks = 2, charge_cap_precursor: charges up to the precursor charge, at most 3, and sum_intensity). The member defaults are the behaviour before these parameters existed (envelopes of at least three peaks, charges 1-3 in every spectrum, the monoisotopic peak keeps its own intensity); the preprocessSpectra_() overload without settings uses them.
| struct OpenMS::ProSEAlgorithm::LocalFragmentEvidence_ |
| struct OpenMS::ProSEAlgorithm::MergedResult |
The pooled evaluation of a multi-file search (the ProSE tool's -out_merged).
| Class Members | ||
|---|---|---|
| PeptideIdentificationList | peptide_ids | PSMs of all files; meta value 'id_merge_index' = input position of the file. |
| bool | protein_fdr_applied = false | picked-protein FDR applied and its filter at FDR:protein (and, with remove_decoys, the decoys removed) |
| vector< ProteinIdentification > | protein_ids |
one pooled run; empty if no file succeeded |
| struct OpenMS::ProSEAlgorithm::MultiFileSearchResult |
Multi-file search result bundle.
Returned by the file-list searchWithModificationAnalysis() overloads. Holds one SearchResult per input file (in per_file, in input order) and a single aggregate result whose peptide_ids are the concatenation of all per-file PSMs and whose modification_analysis is computed once on the pooled set of PSMs.
Special cases for aggregate:
aggregate is left almost-empty (only is_open_search and exit_code are set) — the single-file pooled aggregate would just duplicate per_file[0] and re-run modification analysis on the same PSMs. Callers should use per_file[0] for the result in this case.aggregate.exit_code is set to the first non-OK per-file exit code (so callers can inspect it without walking the per_file vector).build_pooled_aggregate = false, the pooled PSM copy is not retained: aggregate is left almost-empty (only is_open_search and exit_code), exactly as in the single-file case. In OPEN search the pooling still happens internally because the aggregate modification analysis needs it, so aggregate.modification_analysis is populated as usual and only aggregate.protein_ids / aggregate.peptide_ids are released afterwards. In closed search nothing is pooled at all.per_file stay as they are.The aggregate's protein_ids template is taken from the first successful per-file result (search parameters are identical across files by construction), with the primary MS run path overwritten to list every input file. Its PSMs carry the meta value 'id_merge_index', the input position of their file (see poolSearchedFiles_()).
| Class Members | ||
|---|---|---|
| SearchResult | aggregate | |
| bool | decoy_is_prefix = true |
Position of decoy_string (true = prefix, false = suffix). |
| string | decoy_string |
Effective decoy marker resolved from the shared database, for a caller-side merged-PSM protein-FDR step (e.g. the ProSE TOPP tool's -out_merged path). Empty when the search was target-only (decoys=ignore). |
| bool | have_decoys = false | True when the searched databases contained decoys (FDR possible). |
| vector< SearchResult > | per_file | |
| SharedSearchStats | shared |
Configuration / database / fragment-index facts shared across all input files (the index is built once and reused), for the end-of-search report. |
| struct OpenMS::ProSEAlgorithm::PercolatorRescoring |
Outcome of the Percolator rescoring of one search run (see rescoreWithPercolator() and the parameter 'rescore').
With 'rescore' off nothing is attempted: applied is false and skipped_reason empty.
| Class Members | ||
|---|---|---|
| bool | applied = false | True if the PSMs were rescored. |
| string | model |
With 'rescore_reuse:mode' other than per_file: the models that scored the PSMs, as a phrase to show to the user (empty: trained on the run itself) |
| string | skipped_reason |
Why the PSMs were not rescored, as a sentence to show to the user (empty if applied or not attempted) |
| string | warning |
A caveat of an applied rescoring, as a sentence to show to the user (empty if none): Percolator's sanity check replaced the learned model by the best single feature (Percolator::usedDefaultDirection()) |
| struct OpenMS::ProSEAlgorithm::PercolatorTraining |
Options of Percolator's training for the rescoring (the parameters 'rescoring:...'; the defaults are ProSE's training: all PSMs, 10 iterations)
| Class Members | ||
|---|---|---|
| bool | calc_mass_feature = false | Percolator trains on CalcMass ('rescoring:calc_mass_feature'; off, as the tool's default and OpenMS develop since #10717) |
| double | curve_min_gain = 0.002 | learning_curve: relative gain below which a fold stops |
| int | curve_start = 100000 | learning_curve: rows of the first sample |
| bool | fixed_enzyme_termini = false | protein termini are enzymatic sites for enzN/enzC ('rescoring:fixed_enzyme_termini'; UNSAFE: breaks FDR control with Met clipping, keep false) |
| int | iterations = 10 | iterations of the cross-validation training |
| bool | peptide_level = true | also Percolator's peptide-level q-values and PEPs (parameter 'rescoring:peptide_level', default) |
| bool | single_cneg = false | one Cneg candidate instead of three |
| string | solver = "newton" | Percolator's SVM solver (parameter 'rescore_solver', default 'newton') |
| int | subset_max_train = 0 | train on at most this many PSMs (drawn by spectrum, then all are scored; 0 = all) |
| string | subset_mode = "out_of_fold" | with a cap: Percolator's 'subset_mode' ("out_of_fold" | "out_of_fold_avg" | "in_sample" | "learning_curve") |
| double | tron_epsilon = 1e-7 | stopping tolerance of the SVM solver (TRON) |
| string | use_pi0 = "true" | pi0 estimate in the q-values and PEPs ('rescoring:use_pi0': true, false (the executable with TDC), auto) |
| bool | warm_start = false | SVM trainings start from the previous iteration's weights. |
| struct OpenMS::ProSEAlgorithm::ReuseRun_ |
rescore_reuse: a run that trains models: its index in the search (the random halves depend on it) and its PSMs
| Class Members | ||
|---|---|---|
| Size | file_index | |
| PeptideIdentificationList * | peptide_ids | |
| const vector< ProteinIdentification > * | protein_ids | |
| struct OpenMS::ProSEAlgorithm::RunStatistics |
Per-run identification statistics for the end-of-search report.
Populated by collectRunStatistics_() once a single spectrum file has been searched (post-FDR), plus a few fields captured at well-defined points during search() (target/decoy counts pre-FDR, achieved q-value, timing). All counts refer to one input file. Cross-file/shared facts (database, fragment index, configuration) live in SharedSearchStats instead.
| Class Members | ||
|---|---|---|
| double | achieved_psm_fdr = -1.0 | max retained q-value after FDR (<0 = n/a) |
| map< Int, Size > | charge_histogram | precursor charge -> PSM count |
| Size | charge_out_of_range_spectra = 0 | MS2 spectra not searched: their charge is outside precursor:min/max_charge. |
| Size | decoy_psms = 0 | decoy PSMs in the final IDs (after FDR, if applied) |
| bool | fdr_applied = false | true if PSM-level FDR filtering ran |
| double | frag_err_mad = 0.0 | |
| double | frag_err_median = 0.0 | |
| double | frag_err_recommended = 0.0 | |
| bool | frag_tol_valid = false |
true if fragment-error estimate present The same of the annotated b and y fragment peaks (without neutral losses) of these PSMs; within the searched fragment tolerance (unmatched peaks have no error) |
| double | hyperscore_max = 0.0 | |
| double | hyperscore_median = 0.0 | |
| double | hyperscore_min = 0.0 | |
| string | input_file | spectrum file this run searched (basename or path) |
| Size | matched_spectra = 0 | spectra with >=1 retained PSM in the final IDs (after FDR, if applied) |
| map< Size, Size > | missed_cleavage_histogram | missed cleavages -> PSM count |
| Size | ms2_spectra = 0 | number of MS2 spectra in the input |
| double | prec_err_mad = 0.0 | |
| double | prec_err_median = 0.0 | |
| double | prec_err_recommended = 0.0 | |
| bool | prec_tol_valid = false |
true if precursor-error estimate present Mass error estimate (ppm) of the top hits of the target PSMs (at least 10): the median of the signed errors (observed minus theoretical; of the precursor corrected by the hit's isotope error, in an open search of the PSMs without a delta mass, within 0.05 Da of zero, only), their median absolute deviation from it (unscaled), and the recommended symmetric tolerance ceil(|median| + 3 * 1.4826 * MAD) |
| Size | profile_spectra = 0 | MS2 spectra recorded as profile (not centroided) spectra. |
| bool | score_stats_valid = false | true if hyperscore_* below are meaningful |
| double | seconds_calibration = 0.0 | calibration pass wall time (0 if disabled) |
| double | seconds_fdr = 0.0 | FDR filtering wall time (0 if not applied) |
| double | seconds_search = 0.0 | scoring + post-processing wall time |
| Size | target_psms = 0 | target PSMs in the final IDs (after FDR, if applied) |
| Size | unique_peptides = 0 | distinct peptide sequences among top hits |
| Size | unique_proteins = 0 | distinct protein accessions among top hits |
| struct OpenMS::ProSEAlgorithm::SearchContext |
Prepared per-database state shared across multiple spectrum files.
Holds the (decoy-augmented) protein database and the built FragmentIndex so that searching N spectrum files against the same FASTA pays the index build cost only once. Construct via prepareContext() and pass to the context-taking search() overload.
| Class Members | ||
|---|---|---|
| vector< FASTAEntry > | db | |
| bool | decoy_is_prefix = true |
Position of decoy_string (true = prefix, false = suffix). |
| string | decoy_string |
Effective decoy marker carried by |
| bool | electron_ions = false |
True when |
| FragmentIndex | fragment_index | |
| bool | have_decoys = false |
True when |
| bool | release_fragment_index_after_scoring = false |
When true, the context-taking search() overload will release |
| struct OpenMS::ProSEAlgorithm::SearchResult |
Comprehensive search result including modification analysis.
This structure contains all outputs from an open search including:
| Class Members | ||
|---|---|---|
| ExitCodes | exit_code = ExitCodes::EXECUTION_OK | |
| PeptideIdentificationList | inference_only_peptide_ids |
The PSMs whose q-values pass FDR:protein_inference_PSM but not FDR:PSM (targets and decoys, with the q-value as main score): the protein inference and the picked-protein FDR use them with |
| bool | is_open_search = false | |
| OpenSearchAnalysisResult | modification_analysis | |
| PeptideIdentificationList | peptide_ids | |
| PercolatorRescoring | percolator | Outcome of the Percolator rescoring of this run (parameter 'rescore') |
| PeptideIdentificationList | pre_fdr_peptide_ids |
the PSMs before rescoring and FDR:PSM; only filled with setKeepPreFdrPsms(true) |
| vector< ProteinIdentification > | protein_ids | |
| RunStatistics | stats | |
| struct OpenMS::ProSEAlgorithm::SharedSearchStats |
Configuration, database and fragment-index facts shared across all input files of one ProSE invocation.
Computed once (the fragment index is built once and reused), so these costs/counts must NOT be summed per file. Populated by the multi-file searchWithModificationAnalysis() overloads.
| Class Members | ||
|---|---|---|
| bool | calibration_enabled = false | |
| bool | chunked = false | |
| string | database_file | FASTA path (empty for in-memory db) |
| Size | db_decoy_proteins = 0 | decoy entries in the searched (augmented) db |
| Size | db_target_proteins = 0 | target entries in the searched (augmented) db |
| string | decoy_mode | "generated" | "external" | "none (target-only)" |
| string | enzyme | |
| vector< string > | fixed_mods | |
| double | fragment_tol = 0.0 | |
| string | fragment_tol_unit | |
| Size | indexed_fragments = 0 | theoretical fragments in the index (summed over chunks) |
| Size | indexed_peptides = 0 | peptides in the fragment index (summed over chunks) |
| vector< string > | ion_series | |
| Int | max_charge = 0 | |
| Int | min_charge = 0 | |
| Size | missed_cleavages = 0 | |
| bool | open_search = false | |
| bool | peptide_mass_mode = false | |
| double | precursor_tol_lower = 0.0 | |
| string | precursor_tol_unit | |
| double | precursor_tol_upper = 0.0 | |
| double | protein_fdr_threshold = 0.0 | |
| double | psm_fdr_threshold = 0.0 | |
| double | seconds_index_build = 0.0 | decoy generation + fragment index build wall time |
| double | seconds_output = 0.0 | of seconds_total, the wall time after the search of the files: the outputs (set by the caller) |
| double | seconds_qpx_extended = 0.0 | of seconds_output, ProSE's -qpx_extended work: the spectra and predictions of the PSMs (set by the caller) |
| double | seconds_total = 0.0 | whole-search wall time (set by the caller) |
| vector< string > | variable_mods | |
| struct OpenMS::ProSEAlgorithm::SpectraCounts_ |
The MS2 spectra of a spectrum file, those recorded as profile spectra and those that are not searched because their precursor charge lies outside of precursor:min_charge..max_charge (FragmentIndex::precursorCharges())
| Class Members | ||
|---|---|---|
| Size | charge_out_of_range = 0 | |
| Size | ms2 = 0 | |
| Size | profile = 0 | |
| using FileResultConsumer = std::function<void(Size, const SearchResult&)> |
The consumer of the files' results (setFileResultConsumer()): file index, result. Must only read the result.
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| ProSEAlgorithm | ( | ) |
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Helper: does accession carry the decoy marker at the given position? Empty marker → false. Pure std::string (no String dependency).
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The fragment annotations that postProcessHits_() leaves out with defer_peak_annotations, for the hits of peptide_ids (by their "scan_index" in exp), computed as postProcessHits_() computes them.
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Learn per-run fragment ion likelihoods from confident PSMs and annotate every hit with them.
Implements annotate:self_trained_ion_priors. The spectra are split into two folds by the parity of their scan_index. Each fold trains its own FragmentIonLikelihoodModel on its rank-one target hits at target-decoy competition q <= annotate:ion_prior_train_fdr of the native score, computed within the fold, with their reversed sequences matched against the same spectra as noise. Every hit is then scored by the model of the other fold (cross-fitting), so neither a PSM's own spectrum nor its label, nor any other label of its fold, enters the model that scores it; targets and decoys are scored alike. Both folds need annotate:ion_prior_min_psms training PSMs (a fold without a decoy hit has the estimate (0 + 1) / T); otherwise all features of the file are 0 and a warning is logged. This run-level gate is the one place where the labels of a fold reach its own features: at the boundary, a single label (in either fold) decides whether both folds get their cross-fitted features or all zeros. The gate treats targets and decoys alike, but it is not formally independent of the labels. Adds the feature names to the search parameters' extra_features and records the settings that determine the features (annotate:ion_prior_*) and the training set (ion_prior:*). Native scores and the retained candidates are unchanged. Call after PeptideIndexing (target/decoy labels) and before FDR (which overwrites the native scores). Does nothing when the ion priors are off, which includes target-only searches (decoys=ignore).
| [in] | spectra | Preprocessed spectra, indexed by the scan_index meta value of the PSMs (precursor activation selects the ion series). |
| [in] | evidence | Peak lists of the same spectra from preprocessSpectra_. |
| [in,out] | protein_ids | Search parameters of protein_ids[0] receive the feature names and training facts. |
| [in,out] | peptide_ids | Every hit receives the meta values of annotate:ion_prior_features (ion_prior_llr, ion_prior_explained, ion_prior_topk_observed). |
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Adds PeptDeep-prediction-derived rescoring features to the PSMs.
No-op unless both peptdeep:ms2_model and peptdeep:rt_model are set. Warns and does nothing when OpenMS was built without ONNX support, so an ini file that requests the models still runs (without those features) on such a build.
| void annotatePsmQValues | ( | PeptideIdentificationList & | peptide_ids | ) | const |
Annotate PSM q-values by target-decoy competition of the best hit of each spectrum, as FDR:PSM does.
Uses FalseDiscoveryRate (decoy hits are kept and annotated) and replaces hit scores by q-values; no filtering. With FDR:PSM_groups = 'scored_charges' and multiply charged fragments scored (scoring:fragment_charges), PSMs whose best hits were scored with different numbers of fragment charges compete separately, because HyperScore grows with the number of theoretical ions. With one group, or a group without target or decoy hits, all PSMs compete together.
| [in,out] | peptide_ids | PSMs with target_decoy annotations (PeptideIndexing). |
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Finalize protein-level FDR on a COMPLETE protein set (a single input file, or a merged cross-file aggregate).
Runs protein inference (BasicProteinInferenceAlgorithm, see inferProteins()), picked-protein FDR (Savitski et al. 2015), threshold filtering at protein_fdr, then removes decoys and repairs indistinguishable-protein / protein-group / peptide-evidence references so the result stores as schema-valid, decoy-free idXML.
The inference and the picked-protein FDR use the reported PSMs (peptide_ids, e.g. those that pass FDR:PSM) together with inference_only_psms (e.g. those up to FDR:protein_inference_PSM), as ProteomicsLFQ infers proteins on PSMs of a looser cutoff before it filters them at its PSM FDR: more decoy proteins for the picked FDR, and the score of a protein is the best PEP of its PSMs (the raw search engine score without a PEP, see inferProteins()). The proteins and the indistinguishable groups are filtered at protein_fdr, each by its own q-value (the picked FDR of the groups counts a group once, however many targets it holds, so a group can be above the cutoff while its proteins are not), and only the proteins of the groups that pass stay. The reported PSMs stay as they are, but for the decoys that the finalization removes and the PSMs all of whose proteins the filter removed (as ProteomicsLFQ removes them: no reported PSM is left without a protein); the proteins and groups that none of them references afterwards (the decoys, and the proteins only inference_only_psms support) are removed.
Shared by the file-based single-file search() above, mergeFileResults() and the ProSE TOPP tool's single-file finalization, so the exact IDFilter sequence and ordering live in one place (they previously existed as copies that could drift).
Precondition: protein_ids is non-empty and the set is statistically complete — picked-protein FDR does not compose across runs. The caller gates on "decoys present" and "FDR requested"; this helper additionally skips (with a warning) when no decoy proteins survive inference, to avoid target-only q-values.
With remove_decoys false the decoys are kept, as mergeFileResults() keeps them: the picked-protein FDR keeps the decoy proteins, with q-values, the filter at protein_fdr applies to them as to the targets, the decoy PSMs stay, and only the references to the proteins the filter removed go.
| [in,out] | protein_ids | Protein identifications (operates on protein_ids[0]). |
| [in,out] | peptide_ids | The reported PSMs; only the best hit of each is kept, decoy PSMs are removed (unless remove_decoys is false), and so are the PSMs whose proteins all fail the protein FDR. |
| [in] | decoy_string | Accession marker identifying decoy proteins. |
| [in] | decoy_is_prefix | Whether decoy_string is a prefix (true) or suffix (false). |
| [in] | protein_fdr | Picked-protein q-value threshold (expected > 0). |
| [in] | remove_decoys | Remove the decoy PSMs and proteins (the finalization, as in the ProSE tool). |
| [in] | inference_only_psms | PSMs that the inference and the picked-protein FDR use as well, but that are not reported (SearchResult::inference_only_peptide_ids); their identifiers are ignored. No default, so that a caller cannot drop the searches' inference-only PSMs unnoticed: pass the search result's (empty with FDR:PSM 0 or FDR:protein_inference_PSM at or below FDR:PSM), or an empty list for PSMs without them. |
decoy_string, or no decoy protein left after the inference, which uses the best hit of each PSM only). | bool applyDeferredPsmFDR | ( | const std::vector< ProteinIdentification > & | protein_ids, |
| PeptideIdentificationList & | peptide_ids, | ||
| bool | rescored, | ||
| double | psm_fdr, | ||
| PeptideIdentificationList * | inference_only_psms = nullptr |
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| ) | const |
Apply the PSM-level FDR to PSMs that were rescored separately, e.g. with rescoreWithPercolator() on the result of a search with 'rescore' and the FDR filters off.
For PSMs that rescoreWithPercolator() rescored (rescored), the scores are Percolator q-values already. Otherwise they get q-values by annotatePsmQValues() first (FDR:PSM_groups of this instance; without a decoy PSM the conservative 1/T), provided the run has decoy proteins (protein hits with 'target_decoy' "decoy"). The PSMs are then filtered at psm_fdr (IDFilter::filterHitsByScore); target and decoy PSMs are filtered alike, decoys are not removed (that is the protein FDR's finalization, see applyCompleteSetProteinFDR()).
The searches of this class do not call it: their own PSM-level FDR (after the rescoring with 'rescore') takes the decoys of the searched database, so that a run without a decoy PSM is filtered too, with q-values 1/T. Here the run's protein hits are all that tells whether the search had decoys.
| [in] | protein_ids | The runs of the search; the first one tells whether there are decoys. |
| [in,out] | peptide_ids | PSMs of the run, rescored by rescoreWithPercolator() or with their search engine scores. |
| [in] | rescored | Whether rescoreWithPercolator() rescored peptide_ids (its applied). |
| [in] | psm_fdr | PSM q-value threshold. |
| [out] | inference_only_psms | If given, receives the PSMs that the filter removes but whose q-values pass FDR:protein_inference_PSM of this instance, for the protein inference (SearchResult::inference_only_peptide_ids); left empty if nothing was filtered. |
psm_fdr <= 0, no runs or no PSMs, or PSMs that were not rescored without decoy proteins to estimate the FDR with.
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Writes what PeptideIndexing::run(db, protein_ids, peptide_ids) with indexer_parameters writes: the evidences, target_decoy and protein_references of every hit, the protein hits and the PeptideIndexer:* search parameters.
fallback_reason set and nothing changed, if mapping cannot reproduce PeptideIndexing for these identifications (e.g. a hit whose sequence it does not hold); then PeptideIndexing has to run.
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Helper: the PSM-level FDR of one search run, the last step of every search path. With 'rescore' on, rescores the PSMs with Percolator first (rescoreWithPercolator()). Then applies FDR:PSM to Percolator's q-values, or to target-decoy q-values of the search engine scores (annotatePsmQValues(); if has_decoys, also without a decoy PSM). The proteins of the run stay as PeptideIndexing produced them (the protein FDR reduces them to those of the reported PSMs, applyCompleteSetProteinFDR()). Fills the FDR fields of stats. If pre_fdr_psms is given, it receives the PSMs before the rescoring and FDR:PSM (see setKeepPreFdrPsms()), with report:top_hits hits per spectrum; Percolator rescores rescoring:top_hits hits per spectrum (see searchTopHits_()); inference_only_psms receives the PSMs between FDR:PSM and FDR:protein_inference_PSM (with FDR:protein only, see SearchResult::inference_only_peptide_ids; cleared otherwise).
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FDR:PSM on the PSMs of a run after its rescoring (see applyPsmFDR_()); the proteins of the run stay as the search produced them. A rescored run records rescoring:top_hits (if not 0) as 'Percolator:top_hits', a run that was not rescored keeps report:top_hits hits per spectrum. inference_only_psms (if not null) receives the PSMs between FDR:PSM and FDR:protein_inference_PSM.
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Build a strided protein sample for chunked calibration.
Sample size is tied to database_chunk_size_ so the calibration FI never exceeds the user's declared memory budget. Crucial for immunopeptidomics (non-specific digestion) where a fixed-size 5000-protein sample would generate tens of GB of fragment index. Strided rather than first-N so small chunk_size values don't starve the calibration pool (#9182).
Note: very small chunk_size may still produce a calibration pool below calibration_min_psms_; calibration then no-ops silently (follow-up issue: pool PSMs across first-K chunks of the main search loop).
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Build the searched database according to strategy.
Optionally strips pre-existing decoys and/or generates fresh decoys by reversing the target proteins. Produces FASTA entries only — does not construct a FragmentIndex.
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As above, but takes the entries of fasta_db over instead of copying them.
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Sets params on the fragment index of ctx and builds it from ctx.db; with searched_spectra and calibration, restricted to the peptides in the precursor windows of the calibration pass (calibrationPassWindow_())
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Collects the protein occurrences of the sequences of the index peptides candidates.
| [in] | index | The searched fragment index (built from db); must still hold its peptides |
| [in] | db | The searched database |
| [in] | candidates | Indices into FragmentIndex::getPeptides(): the candidates of the hits (in any order, repeats allowed) |
| [in] | indexer_parameters | The parameters PeptideIndexing would run with |
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The window the calibration pass searches with, from the configured one (ppm or Da)
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Helper: capture statistics that must be read BEFORE FDR filtering, namely target/decoy PSM counts (via the "target_decoy" meta set by PeptideIndexing; "target+decoy" counts as target, matching OpenMS FDR semantics) and the HyperScore distribution (FDR overwrites each hit's score with its q-value). Fills stats.target_psms, stats.decoy_psms, stats.hyperscore_* and stats.score_stats_valid.
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Whether co-isolated precursors are searched (chimeric:enabled, which database:chunk_size > 0 does not support). The file-based searches then also read the survey scans.
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Helper: copies of the PSMs of peptide_ids whose q-values (main scores) are above psm_fdr and at most FDR:protein_inference_PSM, appended to inference_only_psms (nothing if that cutoff is not above psm_fdr).
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Helper: fill a RunStatistics with identification counts, distributions and per-run tolerance estimation for one searched file (post-FDR). Silent; rendering is done separately (ProSE TOPP tool or renderRunSummary()). Fields filled at other points (target/decoy counts, achieved FDR, timing) are left untouched.
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Scalar tolerance passed to OpenSearchModificationAnalysis under asymmetric bounds. Uses the tighter of the two positive magnitudes — semantically correct for UniMod Δmass matching precision. OpenSearchModificationAnalysis internally clamps this at MAX_MOD_MAPPING_TOL_ = 0.02 Da; see spec §7 for rationale.
Zero on one side is a legal one-sided window (e.g., [0, 500] Da = "search only positive mass shifts"). In that case std::min() would collapse to 0, passing a useless zero tolerance into the mod analyzer — masked in ppm mode by the internal clamp, but genuinely broken in Da mode. Fall back to the non-zero side so the mod-matching precision reflects the configured tolerance.
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Helper: the entries of db whose accession carries the decoy marker (accessionHasDecoyMarker_())
Number of spectra that ions:by_activation also scores with c and z+1 ions (0 if it is off)
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Helper: the SpectraCounts_ of spectra.
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Keep the strongest peaks in each non-overlapping 100 Da window, including a short final window.
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rescore_reuse joint/static: trains the models on the files of deferred (indices into mfres.per_file, searched with reuse_defer_), then rescores them and applies FDR:PSM
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ProSE parameters made safe to hand to a FragmentIndex.
FragmentIndex declares its own (unused) boolean "decoys" flag with restrictions {true,false}. ProSE's "decoys" parameter is the richer auto/generate/ignore enum and manages decoys at the database level, so forwarding it verbatim would trip FragmentIndex's validation. This returns getParameters() with "decoys" overridden to "false".
| [in] | electron_ions | Also index c and z+1 ions, for electron-activated spectra (ions:by_activation). |
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Helper: whether a PSM of ids has a hit.
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The protein inference of the protein FDR (applyCompleteSetProteinFDR()) and of mergeFileResults().
Runs BasicProteinInferenceAlgorithm on the best hits of peptide_ids and inference_only_psms (copies that hold what the inference reads; the PSMs themselves stay unchanged) for the run protein_ids[0], with the algorithm's 'score_type': "PEP" if every PSM carries a posterior error probability (after Percolator rescoring the main score is the q-value, and the PEP is the meta value MS:1001493), so that a protein scores with the best PEP of its PSMs (as a posterior probability, 1 - PEP), as in ProteomicsLFQ. Otherwise "RAW", ProSE's score 'ln(hyperscore)' if every PSM carries it (as main score, or as a meta value after rescoring or FDR:PSM; e.g. a pooled set of which Percolator rescored only some files), else another raw search engine score that all PSMs carry, or the main score if they carry none.
| [in,out] | protein_ids | Protein identifications; protein_ids[0] gets the scores and the indistinguishable groups, and loses the proteins without PSMs. |
| [in] | peptide_ids | PSMs of the run. |
| [in] | inference_only_psms | Further PSMs of the run (their identifiers are ignored). |
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True if the precursor of spectrum was activated by electrons (ETD, ECD, EThcD or ETciD)
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Helper function to determine if open search should be used based on tolerance.
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Whether a search keeps the PSMs of each file before the rescoring (SearchResult::pre_fdr_peptide_ids)
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Helper: keeps the first n hits of each spectrum (the top n of the search engine score, in the order of postProcessHits_()); changes nothing if no spectrum has more.
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The MS2 spectra of a spectrum file as the searches of this class read them.
Sorted by RT, their peaks by m/z. An mzML file is read in chunks that the OpenMP threads of this thread (at most 16) parse in parallel, in one pass over the file; other files (Bruker .d, Thermo .raw), or an mzML file whose layout the chunks do not support (e.g. compressed), with FileHandler::loadExperiment(). For a second look at the spectra of a search, e.g. those of its PSMs (ProSE's -qpx_extended).
| [in] | filename | The spectrum file (mzML, mzPeak, Bruker .d or Thermo .raw) |
| [in] | keep | If set, only the spectra it accepts are kept, so that the result holds only those in memory. It is called on several threads at once. |
| Exception::BaseException | as FileHandler::loadExperiment() if the file cannot be read |
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Compute local fragment-match evidence on the spectrum before window/top-N peak filtering.
Matches each intact theoretical ion to its nearest experimental peak. Surprise is sum(max(0, -log(2 * tolerance_Da * local_density))). Competition evidence is sum(1 / (1 + alternative_assignments + local_density)). Alternative hypotheses include all configured intact ions and generic H2O/NH3 losses, divided by the fragment charge. Losses only discount ambiguous matches; they are not scored as additional matches and do not require sequence-specific loss eligibility.
| [in] | spectrum | Sorted experimental spectrum after zero-intensity removal and optional deisotoping, before window/top-N peak filtering. |
| [in] | theoretical | Sorted intact ions, with the TSG's aligned integer charge array. |
| [in] | densities | Result of localPeakDensities_(spectrum), reused for all candidates. |
| [in] | tolerance | Positive, finite matching tolerance. |
| [in] | ppm | Whether tolerance is given in ppm rather than Da. |
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localFragmentEvidence_() with the charges of the peaks of theoretical given (charges, one per peak)
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localFragmentEvidence_() restricted to the ions of theoretical with charge <= max_charge, with a caller-owned buffer for the sorted alternative assignments (reused across candidates).
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Peak counts per Da in a fixed +/-50 m/z window around each peak of a sorted spectrum.
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Helper: marks protein_ids as post-processed by Percolator (with parameters pp and the options training), as PercolatorAdapter does; with perc (its last training: capped-training details, RESET) and reuse (rescore_reuse), what they did
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Helper: maximum top-hit score among retained PSMs. After PSM-FDR the score is the q-value, so this is the achieved FDR of the filtered set (-1.0 if empty).
| MergedResult mergeFileResults | ( | MultiFileSearchResult & | result, |
| const StringList & | in_spectra_files, | ||
| bool | remove_decoys = true |
||
| ) | const |
Pools result (moving its PSMs out), infers proteins across files, applies FDR:protein of this instance with picked-protein FDR if decoy proteins were identified and then, by default, removes the decoys.
The evaluation of the ProSE tool's -out_merged, for the result of a multi-file searchWithModificationAnalysis() with the parameters of this instance:
build_pooled_aggregate) is taken as it is. Without one (e.g. build_pooled_aggregate false, or a search of a single file, which builds none), the runs of the files that were searched (exit code EXECUTION_OK) are merged with IDMergerAlgorithm: the PSMs of the files are moved, their runs are copied and stay with the files. 'id_merge_index' is the input position of a PSM's file (its index in in_spectra_files, see poolSearchedFiles_()). The pooled run lists in_spectra_files as its primary MS run paths and gets the current date.per_file[i].inference_only_peptide_ids, which are moved out), with picked-protein FDR (Savitski et al. 2015) and the filter at FDR:protein, provided the inference identified decoy proteins (otherwise a warning, and nothing is filtered). The finalization then removes the decoys, the proteins without a pooled PSM and the references to removed proteins. With remove_decoys false the picked-protein FDR keeps the decoy proteins in the run, with q-values (FalseDiscoveryRate's add_decoy_proteins; the q-values of the targets are the same), and the filter applies to them as to the targets; the decoy PSMs and the decoy proteins that pass are kept, and only the references to the proteins the filter removed go (peptide evidences, indistinguishable proteins and protein groups), so that the result can still be stored, e.g. as idXML. Without the protein FDR the result keeps the targets and decoys.| [in,out] | result | The search result. Its pooled aggregate's identifications are moved into the returned result; without an aggregate, the PSMs of the files that were searched are, while the runs of the files (per_file[i].protein_ids: search parameters, meta values such as PeptDeepRescoring::NCE_META_VALUE, primary MS run paths) stay as they are. The statistics, exit codes, Percolator outcomes and modification analyses stay, too. Unchanged if Exception::IllegalArgument is thrown. If the evaluation fails after the PSMs were moved out (an exception, which is passed on), the files get them back, in their order, as far as the evaluation changed them (at most only the best hit of each, and q-values as above); their PSMs between FDR:PSM and FDR:protein_inference_PSM are gone. |
| [in] | in_spectra_files | The input files of the search, in its order: one per entry of result.per_file. |
| [in] | remove_decoys | Whether the finalization of the protein FDR removes the decoy PSMs and proteins (default, as in the ProSE tool's -out_merged). False keeps those that pass, e.g. to show them; protein_fdr_applied of the result still tells whether the FDR was applied. No effect without the protein FDR. |
| Exception::IllegalArgument | if in_spectra_files has not one entry per file of result. |
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Helper: name as the user sets the parameter (setParameterNamePrefix()), for messages.
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The name of enzyme as PercolatorInfile computes the enzN/enzC/enzInt features of a .pin.
PercolatorInfile knows only Percolator's lower-case enzyme names and treats every other name as "every site is enzymatic". This maps OpenMS's names (e.g. Trypsin/P to trypsinp, PepsinA to pepsin) and returns "no_enzyme" for an enzyme Percolator does not know. rescoreWithPercolator() and the ProSE tool's -out_pin use it.
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The enzyme argument of PercolatorInfile (store(), the in-process rescoring) for the enzN/enzC/enzInt features of the search's enzyme: its Percolator name (pinEnzymeName(); a ProteaseDB enzyme without one keeps its name, so that PercolatorInfile uses its cleavage rule) with "+termini" (fixed_enzyme_termini: protein termini and the position after an initial Met are cleavage sites) or "+keep_termini" (the features of ProSE int15, the default).
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Helper: pool the runs of the files of per_file that were searched (exit code EXECUTION_OK) with IDMergerAlgorithm into one run (protein_ids, one entry) that lists in_spectra_files as primary MS run paths, and their PSMs (peptide_ids). Copies the identifications, or with move_psms moves the PSMs out of per_file (leaving them empty) and copies the runs, which stay with the files. 'id_merge_index' of a PSM is the input position of its file (its index in per_file and in_spectra_files, per occurrence: the same file given twice has two), also when other files failed. Shared by the multi-file search's pooled aggregate and mergeFileResults().
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Filter and annotate search results.
Trims per-spectrum candidate hits to the top N and converts them into PeptideIdentification objects, adding requested PSM annotations and populating protein-level search metadata.
| [in] | exp | Input MS experiment providing spectra/metadata for annotation. |
| [in,out] | annotated_hits | Per-spectrum candidate hits (trimmed to top_hits in-place). |
| [in] | pool_stats | Per-spectrum summary of the full candidate pool as collected by scoreSpectraAgainstIndex_(), used for the pool-derived PSM features (delta score, z-score, candidate count). Must match annotated_hits in size. |
| [out] | protein_ids | Output container for protein-level identification and search metadata. |
| [out] | peptide_ids | Output container for spectrum-level peptide identifications (PSMs). |
| [in] | top_hits | Number of top-scoring hits to retain per spectrum (searchTopHits_()). |
| [in] | modifications_fixed | Fixed modifications (by name) used during the search. |
| [in] | modifications_variable | Variable modifications (by name) used during the search. |
| [in] | peptide_missed_cleavages | Allowed missed cleavages in digestion. |
| [in] | precursor_mass_tolerance | Precursor mass tolerance value. |
| [in] | fragment_mass_tolerance | Fragment mass tolerance value. |
| [in] | precursor_mass_tolerance_unit_ppm | Precursor tolerance unit ("true"->ppm, "false"->Da). |
| [in] | fragment_mass_tolerance_unit_ppm | Fragment tolerance unit ("true"->ppm, "false"->Da). |
| [in] | precursor_min_charge | Minimum precursor charge considered. |
| [in] | precursor_max_charge | Maximum precursor charge considered. |
| [in] | enzyme | Digestion enzyme name. |
| [out] | database_name | Database file name used for the search (stored in protein_ids). |
| [in] | evidence_spectra | Peak lists retained before local/top-N filtering, aligned with exp; required when annotate:local_fragment_evidence is enabled, otherwise unused. |
| [in] | defer_peak_annotations | Leave out the fragment annotations (annotate:PSM fragment_annotation): the caller adds them with annotateFragmentPeaks_() to the hits it keeps. |
| [in] | evidence_parent | For the spectra of exp after the first evidence_spectra->size() (the residual spectra of a chimeric search), the index of the spectrum whose peak list in evidence_spectra they use; else evidence_spectra must be as long as exp. |
| SearchContext prepareContext | ( | const std::vector< FASTAFile::FASTAEntry > & | fasta_db | ) | const |
Build a SearchContext (decoy-augmented database + FragmentIndex) for reuse.
Performs the database preparation and FragmentIndex construction steps so that subsequent calls to search(spectra, ctx, ...) can reuse the same index across many spectrum files. If decoy generation is enabled (parameter "decoys"), decoys are generated and shuffled into the returned context's db member exactly once here.
| [in] | fasta_db | Protein sequence database as FASTA entries. |
Thread-safety: concurrent search() calls reading the same SearchContext are safe (per FragmentIndex query thread-safety contract) as long as calibration:enabled is off. A successful calibration sets the calibrated tolerances as query parameters of the context's FragmentIndex until the call returns, so calibrated searches must not share a context concurrently. Do not call prepareContext() concurrently on the same algorithm instance.
| SearchContext prepareContext | ( | const std::vector< FASTAFile::FASTAEntry > & | fasta_db, |
| bool | electron_ions | ||
| ) | const |
Build a SearchContext whose FragmentIndex also holds c and z+1 ions.
As prepareContext(fasta_db). With electron_ions, the index also holds c and z+1 ions, which ions:by_activation scores for electron-activated spectra (ETD, ECD, EThcD, ETciD). Only those spectra are matched against them, so other spectra get the same candidates as from prepareContext(fasta_db). Use it when the spectra to search contain such spectra: search() otherwise builds a temporary index with these ions for each call.
| [in] | fasta_db | Protein sequence database as FASTA entries. |
| [in] | electron_ions | Also index c and z+1 ions. |
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Helper: preprocessSpectra_() with the settings of the search, after warning about spectra (of input; empty: not named) that cannot be searched as intended: no MS2 spectra (e.g. an MS1-only file), MS2 spectra recorded as profile spectra, MS2 spectra whose precursor charge lies outside of the charge range (not searched). ion_evidence is filled with annotate:self_trained_ion_priors only.
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filter, deisotope, decharge spectra
deisotoping sets the envelope rule of the deisotoper (see DeisotopingSettings_).
With ion_evidence, a compact copy of every spectrum's peaks is kept for the self-trained ion priors (annotate:self_trained_ion_priors): the peaks after deisotoping, before the window and top-N filters, or, with ion_evidence_scored_peaks, the peaks that are scored. The lists are indexed like exp after its sort by RT.
With query_spectra, a copy of each spectrum after zero-intensity removal and m/z sorting, before deisotoping and local filtering, is kept for candidate retrieval (fragment:query_spectrum=raw); with evidence_spectra, a copy after deisotoping, before window/top-N filtering (annotate:local_fragment_evidence). Both are aligned with exp after its sort by RT.
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preprocessSpectra_() with the DeisotopingSettings_ defaults (the behaviour before its parameters existed)
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Free the protein and peptide identifications of result with the OpenMP threads (destroyed one after the other, the hits of a file take tens of ms at the end of a run). Leaves them empty; the statistics are kept.
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Removes the repeated entries of a protein database, which searching would report twice (e.g. a FASTA file concatenated twice, or contaminants added twice).
An entry whose accession (FASTA identifier) an entry before it has, with the same sequence, is removed (the first one stays, with its description); the number removed is logged as a warning that names source.
| [in,out] | db | The database, in file order. |
| [in] | source | The database file (or another name of the database) for the messages. |
| Exception::ParseError | if two entries of one accession have different sequences: they cannot both be reported under it, and which one is meant is unknown. The message names the accession and source. |
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Helper of removeUnreportedProteins() with PSMs besides the reported ones: keeps the protein hits of each run of protein_ids that a PSM of the run (by identifier) in peptide_ids or also_referencing references, as IDFilter::removeUnreferencedProteins() does for one list (the protein groups are not updated)
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Removes the proteins that no PSM references, and them from the protein groups.
Keeps the protein hits of each run of protein_ids whose accession a peptide evidence of a hit of a PSM of the run (by identifier) in peptide_ids or also_referencing names (IDFilter::removeUnreferencedProteins() for one list), and updates the indistinguishable groups and the protein groups of each run (IDFilter::updateProteinGroups()). The protein FDR does this after its filters (applyCompleteSetProteinFDR()); a search leaves the proteins of a run as PeptideIndexing produced them (those of all hits, also of the PSMs that FDR:PSM removed).
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Render the modification-discovery section (top PTMs, unknown delta masses) to os. Shared by the per-file and aggregate report blocks.
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Render a human-readable single-run summary block (configuration recap, database/index stats, identification results, tolerance estimate, timing and — for open searches — modification discovery) to os. shared carries the configuration/db/index context (built once).
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Serialize the complete end-of-search report to a machine-readable YAML string: shared configuration/database/index facts, per-file identification statistics, the output-file manifest (label -> written paths) and failed/total file counts. Hand-rolled (no YAML library dependency); every string scalar is double-quoted so values containing ':' (e.g. Windows paths) or other YAML metacharacters round-trip safely, and non-finite numbers are emitted as null.
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The total time of a run, by part: "total <s>s | search of the file(s) <s>s | outputs <s>s
(of which -qpx_extended <s>s)", from sh's seconds_total, seconds_output and seconds_qpx_extended (the last part only if positive). The end of the report of renderRunSummary(), and of ProSE's report of several files.
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prepareContext(fasta_db, electron_ions) that takes the entries of fasta_db over instead of copying them. With searched_spectra, the index holds only the peptides these spectra can reach (see FragmentIndex::build): the context then serves only one search of these spectra, without calibration. decoy_parity_report: logs the candidate counts of ctx's fragment index per label and subclass (no-op otherwise)
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The end of a multi-file search with 'rescore' (with pool_rescoring_), for the files with PSMs that Percolator could not rescore on their own (e.g. fewer than 100 PSMs, or no decoy PSM).
The files that Percolator rescored on their own keep their results. If there are some, a Percolator model is trained (Percolator::trainPSMs()) on their PSMs before the rescoring (SearchResult::pre_fdr_peptide_ids), at most at most 'percolator:max_train_psms' / (the input files) of each (transferQuota_(), transferSample_(): reproducible random spectra; a file rescored on its own keeps only these after its rescoring, unless setKeepPreFdrPsms()), and scores the other files (Percolator::scorePSMs()): on the scale of its training PSMs (the score a PSM would get among them, their PEP at that score). If Percolator rescored no file on its own, the PSMs of all files are rescored together, with one cross-validated model (rescoreWithPercolator(), with the cap). Back in its file, each PSM gets the q-value of target-decoy competition of the file's PSMs on the model's SVM score, (D + 1) / T as Percolator's own (tdcQValues_(); FDR per run), on which FDR:PSM filters (and FDR:protein_inference_PSM collects); the runs are marked as rescored by Percolator, and percolator.warning of each such file says how it was rescored. If that fails (e.g. fewer than 100 PSMs in all), the files keep their search engine scores (see mergeFileResults()). In any case the copies are released (unless setKeepPreFdrPsms()); the proteins of each file stay as the search produced them.
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The hits per spectrum that Percolator rescores: rescoring:top_hits, or report:top_hits if that is 0.
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Rescore the PSMs of one search run in-process with Percolator, as the parameter 'rescore' does in every search.
Trains Percolator (Percolator::rescorePSMs) on the standard PIN feature set (PercolatorInfile::getStandardFeatureSet) plus the search engine features of the run (the search parameters' 'extra_features', or 'score' without them), with train_best_positive, target-decoy competition (post_processing_tdc), the PSMs for training up to max_train_psms (subset_max_train) and nonparametric PEPs, as the percolator executable. Percolator q-values become the main score (score type "q-value"; the SVM score and PEP are kept as meta values), only the best hit per spectrum is kept, and the runs are marked as post-processed by Percolator (search engine "Percolator", version "3.08-vendored", meta value 'percolator' and the "Percolator:*" search parameter meta values PercolatorAdapter writes).
The searches of this class call it for every run when the parameter 'rescore' is on (the default), before their PSM-level FDR, which then filters on the rescored PSMs' q-values. Call it directly only to rescore PSMs of a search with 'rescore' off, whose FDR filtering must then be off too (FDR:PSM and FDR:protein 0), so that Percolator gets all target and decoy PSMs.
The PSMs are not rescored, and stay unchanged, with fewer than 100 PSMs, without decoy PSMs or if Percolator fails; skipped_reason of the result then says why. Nothing is logged about the skip, so that the caller can report it with its context (e.g. the input file).
| [in,out] | protein_ids | The runs of the search; the first one gives the enzyme and the search engine features. Marked as post-processed by Percolator if the PSMs were rescored. |
| [in,out] | peptide_ids | Target and decoy PSMs of the run (with 'target_decoy' annotations and peptide evidences, as from PeptideIndexing). |
| [in] | threads | Threads Percolator may use (values above 9 use 9, values <= 0 use 3); never more than OpenMP allows. Cross-validation trains 9 models at a time (3 folds x 3 Cneg values), so more than 9 do not help. The results do not depend on it. |
| [in] | max_train_psms | The most PSMs Percolator trains on (its subset_max_train; 0: all, the default); all are scored. The searches pass 'percolator:max_train_psms'. |
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rescoreWithPercolator(); with dropped, the hits it drops are moved there (as memory allows) instead of destroyed; training: the options of Percolator's training (parameters 'rescoring:...' and 'rescore_solver'); extra: further Percolator parameters (rescore_method 'reset': the RESET parameters). With reuse, the PSMs are scored as it says instead of by a model trained on them.
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rescore_reuse: rescores the PSMs of a run (file file_index of the search) with model, as rescoreWithPercolator_(); with check_shift, a run whose PSMs (targets and decoys, labels unused) differ from the training PSMs by more than reuse_max_shift_ is rescored by Percolator trained on it instead
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The residual spectrum of a co-isolated PSM: spectrum without the peaks removed (ascending indices), with precursor as its precursor and without the meta values of CoIsolationDetector::annotate()
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Decide how to obtain/recognise decoys for db.
Detects pre-existing decoys via the common-marker heuristic (OpenMS::DecoyHelper), falling back to the configured decoy_prefix for custom markers, then maps the "decoys" mode (auto/generate/ignore) onto a concrete DecoyStrategy_.
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Whether a single-use context may index only the peptides the searched spectra can reach: not with chimeric search, whose co-isolated precursors the index restriction does not know. With calibration, the restriction covers the windows of the calibration pass (see buildFragmentIndex_()).
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Helper: moves the PSMs of pooled (empty afterwards) to their files of per_file (searchedFileOf_(); others are dropped), without 'id_merge_index', in their order, as PSMs of the files' runs (their identifiers), appended to the files' PSMs.
rescore_reuse: whether spectrum spectrum of file file is drawn as a calibration spectrum (whose decoys set the scale of a model; never trained on) at the calibration fraction fraction: from the 5 % of the spectra whose draw is highest, so never a training spectrum (reuseTrainingDraw()) at any fraction
rescore_reuse: whether spectrum spectrum (its index in the run) of file file is drawn for the training of the models at the sampling fraction fraction (rescore_reuse:max_train)
A seeded hash-uniform draw per spectrum (all PSMs of a spectrum together), independent of its position: a capped training sample is spread at random over the whole run, not its first spectra. The 5 % of the spectra whose draw is highest are calibration spectra and never drawn for training.
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Reversed sequence keeping the C-terminal residue and every residue's modification: the noise hypothesis of a confident PSM.
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Run a fast calibration pass on a subset of spectra to estimate mass accuracy.
Scores a TIC-ranked subset of spectra against the fragment index, collects precursor and fragment mass errors from high-confidence PSMs, and returns calibrated tolerances using median + 3*MAD estimation.
| [in] | spectra | Preprocessed MS/MS spectra (subset is selected internally by TIC). |
| [in,out] | fragment_index | Pre-built fragment index for candidate lookup. |
| [in] | db | Protein database (for sequence reconstruction of candidates). |
| [in] | query_spectra | Optional spectra for candidate retrieval, aligned with spectra. |
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Score all spectra against one FragmentIndex.
Shared by the non-chunked and chunked search paths. Appends per-scan AnnotatedHit_ entries to annotated_hits and prunes each spectrum to max(searchTopHits_(), 2) candidates to bound memory. Expects a pre-built FragmentIndex whose parameters already reflect any calibrated tolerances the caller wants to apply.
| [in] | spectra | Preprocessed MS2 spectra to score. |
| [in] | fi | Pre-built FragmentIndex to query for candidates. |
| [in] | db | Database the FragmentIndex was built from, used to reconstruct candidate sequences. |
| [in] | generators | Generators for the theoretical spectrum of each candidate, chosen per spectrum. |
| [in] | effective_fragment_tol | Fragment mass tolerance to score with (calibrated, if calibration ran). |
| [in] | fragment_mass_tolerance_unit_ppm | Whether effective_fragment_tol is in ppm rather than Da. |
| [in] | open_search_mode | Whether to record the precursor delta mass on each hit. |
| [in,out] | annotated_hits | Per-spectrum candidate hits, one entry per spectrum. |
| [in,out] | pool_stats | Per-spectrum summary of the full (unpruned) candidate pool, one entry per spectrum. Accumulated rather than overwritten, so chunked callers can pass the same vector for every chunk. |
| [in] | progress_label | Label shown by the progress logger for this scoring pass. |
| [in] | query_spectra | Optional spectra for candidate retrieval, aligned with spectra (fragment:query_spectrum=raw); scoring always uses spectra. |
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Fragment charges that score a candidate of the given precursor charge; also used by the calibration pass.
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The order in which scoreSpectraAgainstIndex_() scores the spectra: the scan indices sorted by the neutral precursor mass of the first precursor (of query_spectra if given; an unknown charge counts as 1), ties by scan index; spectra without a finite mass at the end in scan order.
| ExitCodes search | ( | const std::string & | in_spectra, |
| const std::string & | in_db, | ||
| std::vector< ProteinIdentification > & | prot_ids, | ||
| PeptideIdentificationList & | pep_ids | ||
| ) | const |
Search spectra in a spectrum file (mzML or Bruker .d) against a protein database using an FI-backed workflow.
Populates protein and peptide identifications, including search meta data, PSM hits, and search engine annotations. Parameters are taken from this instance (DefaultParamHandler).
| [in] | in_spectra | Input path to the spectra file (mzML or Bruker .d) containing MS/MS spectra to search. |
| [in] | in_db | Input path to the protein sequence database in FASTA format. |
| [out] | prot_ids | Output container receiving search meta data and protein-level information. |
| [out] | pep_ids | Output container receiving spectrum-level peptide identifications (PSMs). |
Side effects:
Errors:
| ExitCodes search | ( | PeakMap & | spectra, |
| const std::vector< FASTAFile::FASTAEntry > & | fasta_db, | ||
| std::vector< ProteinIdentification > & | prot_ids, | ||
| PeptideIdentificationList & | pep_ids | ||
| ) | const |
In-memory search: search spectra against a protein database without file I/O.
Same as the file-based search() but takes pre-loaded spectra and FASTA entries directly. Spectra are preprocessed in-place (filtered, deisotoped, normalized).
| [in,out] | spectra | MS/MS spectra to search (preprocessed in-place). |
| [in] | fasta_db | Protein sequence database as FASTA entries. |
| [out] | prot_ids | Output protein-level identifications. |
| [out] | pep_ids | Output spectrum-level peptide identifications (PSMs). |
Internally this is a thin wrapper around prepareContext() + the context-taking search() overload, so the FragmentIndex is rebuilt on every call. For repeated searches against the same database, prefer calling prepareContext() once and reusing the resulting SearchContext.
| ExitCodes search | ( | PeakMap & | spectra, |
| SearchContext & | ctx, | ||
| std::vector< ProteinIdentification > & | prot_ids, | ||
| PeptideIdentificationList & | pep_ids | ||
| ) | const |
In-memory search using a pre-built SearchContext.
Searches spectra against the database and FragmentIndex held in ctx. The fragment index build cost (decoy generation, peptide/fragment generation, sorting, bucketing) is paid by prepareContext() and is not repeated here, making this overload the right choice when searching many spectrum files against the same database.
| [in,out] | spectra | MS/MS spectra to search (preprocessed in-place). |
| [in,out] | ctx | Pre-built SearchContext from prepareContext(). Taken by non-const reference because the underlying FragmentIndex query API is non-const, even though the index content is not modified during the search; the db member is also handed non-const to the downstream PeptideIndexing step (which requires a non-const reference). |
| [out] | prot_ids | Output protein-level identifications. |
| [out] | pep_ids | Output spectrum-level peptide identifications (PSMs). |
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Chunked database search implementation.
Splits full_db into chunks of database_chunk_size_ proteins, builds a FragmentIndex per chunk, scores all spectra against each chunk, and accumulates hits before a single post-processing pass. full_db is the already-resolved database (decoys generated/stripped per strategy); strategy carries the effective decoy marker for PeptideIndexing and FDR. Taken by non-const reference because PeptideIndexing::run() mutates it.
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Search the precursors co-isolated with the selected one in what the spectrum's other PSMs leave of it (chimeric:enabled), after the andes chimeric cascade.
Runs one round per co-isolated precursor rank: round k searches the k-th co-isolated precursor (meta values of CoIsolationDetector::annotate()) of every spectrum with at least one hit, at its m/z and charge, against the spectrum without the peaks claimed so far. The best hit of the spectrum claims its peaks first, and the best hit of each co-isolated precursor (its winner) claims its peaks before the next round: every peak within the fragment tolerance of one of its theoretical ions. Hits with the best hit's peptide are left out. A co-isolated precursor is dropped when its winner repeats a PSM of the spectrum (the peptide and charge of a hit of the spectrum or of an earlier winner). The residual spectra are scored in batches and not kept.
| spectra | The searched spectra | |
| annotated_hits | The hits of each spectrum of spectra | |
| fi | The fragment index of the search | |
| db | The searched database | |
| generators | Theoretical spectrum generators of the search | |
| fragment_tol | Fragment mass tolerance | |
| fragment_tol_ppm | Whether fragment_tol is in ppm (else Da) | |
| open_search_mode | Whether the search is an open search | |
| [out] | psms | The co-isolated PSMs, in spectrum order and, within a spectrum, in detection order |
| [out] | hits | The hits of each of psms (at most chimeric:top_hits and report:top_hits), the winner first |
| [out] | pool_stats | The candidate-pool statistics of each of psms |
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Helper: whether the file of pf was searched (exit code EXECUTION_OK, with a run)
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Helper: the file of per_file that a pooled PSM id comes from ('id_merge_index', see poolSearchedFiles_()), or -1 if it has none of a file that was searched
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The hits per spectrum that the search keeps (before the rescoring): without 'rescore' (or with decoys 'ignore', which leaves Percolator no decoys) report:top_hits; with it the hits Percolator rescores, and with a copy for a .pin (setKeepPreFdrPsms()) also the reported ones.
| SearchResult searchWithModificationAnalysis | ( | const std::string & | in_spectra, |
| const std::string & | in_db, | ||
| const std::string & | output_base_name = "" |
||
| ) | const |
Search with comprehensive results including modification analysis tables.
This method performs a peptide database search and additionally returns structured modification analysis results for open search mode. This is the recommended method for modification discovery workflows.
The method automatically:
| in_spectra | Input path to the spectra file (mzML or Bruker .d) containing MS/MS spectra |
| in_db | Input path to the protein sequence database in FASTA format |
| output_base_name | Optional base name for output files (TSV tables) |
Example usage:
| MultiFileSearchResult searchWithModificationAnalysis | ( | const std::vector< std::string > & | in_spectra_files, |
| const std::string & | in_db, | ||
| const std::vector< std::string > & | output_base_names = {}, |
||
| const std::string & | aggregate_base_name = "", |
||
| bool | build_pooled_aggregate = true |
||
| ) | const |
Multi-file search with modification analysis (FASTA file path).
Convenience overload that loads the FASTA database from in_db and delegates to the in-memory multi-file overload. The database file path is recorded in each per-file ProteinIdentification's SearchParameters (and on the aggregate result).
build_pooled_aggregate is forwarded unchanged; see the in-memory overload for its exact effect on aggregate.
| MultiFileSearchResult searchWithModificationAnalysis | ( | const std::vector< std::string > & | in_spectra_files, |
| const std::vector< FASTAFile::FASTAEntry > & | fasta_db, | ||
| const std::vector< std::string > & | output_base_names = {}, |
||
| const std::string & | aggregate_base_name = "", |
||
| bool | build_pooled_aggregate = true |
||
| ) | const |
Multi-file search with modification analysis (in-memory FASTA).
Builds a single SearchContext (decoy generation + FragmentIndex) from fasta_db and reuses it across all input spectrum files. Each input file produces its own SearchResult including a per-file modification analysis (TSV written if a non-empty per-file base name is provided). An additional aggregate SearchResult is computed by pooling all per-file peptide identifications and running modification analysis once on the pooled set (see build_pooled_aggregate to skip retaining that second copy of every PSM).
Progress: the search of each input file is a progress step of its own, labelled "Searching <file name> (i of n)..." (i from 1, in input order). Consumers identify the per-file step by this label. Every start of a per-file step is balanced by its end, also if the search of the file fails or throws; the steps nested in it are not exception-safe (one that an exception leaves stays open). The steps of the search of the file (index build, filtering, scoring, ...) nest one level below it (depth 0 for the per-file step, depth 1 inside), but this nesting holds only with one OpenMP thread: with more, the background reading of the next mzML file shares the static step depth of the ProgressLogger, and the depths reported are not reliable. Chunk-major searches (database:chunk_size > 0 with a database of more proteins, decoys included, than the chunk size) emit no per-file step.
| [in] | in_spectra_files | Spectrum file paths (mzML or Bruker .d). |
| [in] | fasta_db | Protein sequence database as FASTA entries. |
| [in] | output_base_names | Optional per-file base names for modification-analysis TSV outputs. Must be empty or have the same length as in_spectra_files. Empty entries skip TSV writing for that file. |
| [in] | aggregate_base_name | Optional base name for the aggregate modification-analysis TSV output. Empty disables aggregate TSV writing (the aggregate analysis is still computed). |
| [in] | build_pooled_aggregate | Whether the pooled PSM set is retained in aggregate. Pooling copies every per-file PSM a second time, so callers that never read aggregate.peptide_ids should pass false. With false, aggregate.protein_ids and aggregate.peptide_ids come back empty (only is_open_search and exit_code are set); the aggregate modification analysis is unaffected — in open search the pooled set is still built internally, analyzed, and then released. Default true (unchanged behavior). |
Errors:
output_base_names is non-empty and its size differs from in_spectra_files. | SearchResult searchWithModificationAnalysis | ( | PeakMap & | spectra, |
| const std::vector< FASTAFile::FASTAEntry > & | fasta_db, | ||
| const std::string & | output_base_name = "" |
||
| ) | const |
In-memory search with modification analysis: no file I/O required.
Same as the file-based searchWithModificationAnalysis() but takes pre-loaded data.
| [in,out] | spectra | MS/MS spectra (preprocessed in-place). |
| [in] | fasta_db | Protein sequence database as FASTA entries. |
| [in] | output_base_name | Optional base name for TSV output files. |
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protected |
The multi-file searchWithModificationAnalysis() overloads: searches fasta_db_given, or, with in_db set, the database read from *in_db (while the first spectrum file is read in the background), whose entries the search then takes over instead of copying them.
|
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Lets the multi-file searchWithModificationAnalysis() overloads hand each file's result to consumer as soon as it is final, e.g. to write it while the next file is searched.
Called for every file but the last whose search succeeded, right after its search, unless the search changes the results of the files later: the rescoring is deferred (rescore_reuse 'joint' or 'static', for the files of the training), Percolator could not rescore the file on its own (with 'rescore', see rescoreFilesTogether_()), or the database is chunked. (The pooled aggregate is built from copies, after the consumers.) The results lack what the file-path overload sets after the search (SearchParameters::db of the first ProteinIdentification). The consumer runs on a helper thread with two of the OpenMP threads of the searching thread (one if it has fewer than 8), which the search takes back after the consumer returned, where it starts its next phase; the next file's search goes on meanwhile. Only one consumer runs at a time, and the search waits for the last one before it returns. If the searching thread has fewer threads than the machine has cores (then a read in the background takes threads from it, see BackgroundSpectra in the .cpp) or only one, the consumer runs on the searching thread itself, before the next file. Exceptions of the consumer are ignored: it handles its errors itself. An empty function (default) turns this off.
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With keep = true, the multi-file searchWithModificationAnalysis() overloads also return the PSMs of each file before the rescoring ('rescore') and FDR:PSM, in SearchResult::pre_fdr_peptide_ids (e.g. for a Percolator .pin): the search engine scores, every reported hit and the decoys. Default: false (no copy).
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The prefix under which the caller's user sets the parameters of this instance (e.g. 'Search:' in the ProSE tool): the messages of parameters it refuses name them with it (e.g. Search:precursor:min_charge). Set it before setParameters(). Default: none.
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Generators for scoring, annotation and calibration, configured from the ions:* parameters.
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Makes the PSMs of each file of a multi-file search its part of the pooled result (mergeFileResults()), so that per-file outputs agree with the pooled one.
The pooled finalization of the protein FDR removes PSMs that the files still have: the PSMs all of whose proteins fail the protein FDR and, with its remove_decoys, the decoy PSMs; it also drops the peptide evidences of the proteins that failed. This moves the PSMs of merged (merged.peptide_ids, empty afterwards) into the files of result by their 'id_merge_index' (the input position of their file, which is removed), in their order, as the PSMs of the runs of the files (their identifiers). The proteins of each file become the proteins of the pooled run (merged.protein_ids) that its PSMs reference, with their pooled scores (e.g. the q-values of the protein FDR, with the pooled run's score type and inference engine) and the pooled indistinguishable groups and protein groups of these proteins (with their pooled scores), so that each file's outputs state why its proteins are there. Without a pooled run, the file keeps the proteins its PSMs reference (removeUnreportedProteins()). A file without a PSM in merged gets none. Store the pooled PSMs (e.g. as idXML) before.
| [in,out] | merged | The pooled result of result. Its PSMs are moved out. |
| [in,out] | result | The multi-file search that merged pools. The files that were searched (exit code EXECUTION_OK) get their PSMs from merged; the statistics stay (see updateFinalStats()). |
| Exception::IllegalArgument | (and nothing changed) if a PSM of merged has no 'id_merge_index' of a file of result that was searched. |
|
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rescoring:stratum_q_values: grouped target-decoy competition on the best hit of every spectrum that has the meta value percolator_score; stratum 'protein_nterm' (an evidence at a protein N-terminus, or only Met-clipped evidences: aa_before M at start 1) or 'other'; q = (decoys + 1) / max(targets, 1) in percolator_score order (ties: decoys first), monotone. Sets the meta values 'stratum' and 'stratum_q_value'; a best hit without percolator_score gets only 'stratum_q_value' = 1.
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Helper: target-decoy q-values of the PSMs of peptide_ids by their main scores, competing all together (FalseDiscoveryRate, decoys kept), and without a decoy PSM the conservative 1/T of the T PSMs instead of 0. With plus_one, the +1-corrected (D + 1) / T at every score, as Percolator estimates its q-values, also with decoys (for PSMs that Percolator scored, see rescoreFilesTogether_()).
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rescore_reuse: trains the two cross-fitted models on PSMs of runs (see the parameter); throws if Percolator cannot train them
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The PSMs of a file rescored on its own that the model for the files too small for one of their own may train on (rescoreFilesTogether_()): percolator:max_train_psms / pool_files_ (no limit with max_train_psms 0)
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The spectra (positions in ids) of a seeded random sample of whole spectra with at most max_psms PSMs, counting at most hits_per_spectrum hits of a spectrum; in ascending order. A spectrum's place in the draw depends only on the spectrum (its reference, retention time and precursor m/z), so the sample of a list is also the sample of any of its sublists that contain it (the copy of a file reduced to it after the rescoring, see applyPsmFDR_()), and it is the same on every platform. Spectra without hits are left out.
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Recompute result-level statistics (matched count, unique peptide/protein counts, charge & missed-cleavage histograms, target/decoy counts and achieved PSM FDR) from a FINAL, post-processed PSM list. Use this when the identifications are mutated AFTER search() returns — e.g. the ProSE TOPP tool's Percolator rescoring + deferred FDR — so the report reflects what the user actually receives. Does NOT touch ms2_spectra, the HyperScore distribution (captured pre-FDR during the search) or any timing field. fdr_applied records whether a PSM-level FDR filter was applied; when true achieved_psm_fdr is set to the maximum retained q-value.
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overrideprotectedvirtual |
This method is used to update extra member variables at the end of the setParameters() method.
Also call it at the end of the derived classes' copy constructor and assignment operator.
The default implementation is empty.
Reimplemented from DefaultParamHandler.
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staticprotected |
Helper: makes SEARCH_ENGINE_SCORE the main score of every PSM of ids (IDScoreSwitcherAlgorithm; the former main score stays a meta value), if every PSM with hits has it (as main score or as meta value). Returns whether it did; otherwise ids is unchanged. For a set of PSMs of which Percolator rescored only some (see mergeFileResults()).
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calibration:min_fragment_tolerance (0: by unit)
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calibration:min_precursor_tolerance (0: by unit)
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calibration:precursor_sigma_factor (0: the precursor window is kept)
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calibration:sigma_factor (fragment)
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chimeric:enabled: also search the precursors co-isolated with the selected one
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staticconstexpr |
PSM meta value of a chimeric search (chimeric:enabled): 1 for a PSM of the selected precursor, 2 for one of a precursor co-isolated with it
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0 = disabled; >0 = chunk DB into groups of this many proteins
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decoy_method 'keep_termini'
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decoy_parity_report
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Resolved MS2 deisotoping request (param fragment:deisotope != "false"). preprocessSpectra_ still gates on Deisotoper::isToleranceSupported() so the deisotoper is never called out of range (it would throw -> terminate in the OpenMP region). See OpenMS#9619.
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MS2 deisotoping rule (fragment:deisotope_*)
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FDR:protein_inference_PSM: q-value cutoff of the PSMs of the protein inference.
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FDR:PSM_groups: separate competitions per number of scored fragment charges.
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mutableprotected |
The PSMs of the most recent search(spectra, ctx, ...) that pass FDR:protein_inference_PSM but not FDR:PSM (only with FDR:protein, see SearchResult::inference_only_peptide_ids), bridged to the caller like last_run_stats_.
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annotate:ion_prior_features holds ion_prior_explained
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annotate:ion_prior_features holds ion_prior_llr
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annotate:ion_prior_features holds ion_prior_topk_observed
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annotate:ion_prior_max_fragment_charge (capped at precursor charge - 1)
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annotate:ion_prior_min_psms: training PSMs required per fold
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annotate:ion_prior_model
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annotate:ion_prior_peaks == scored
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annotate:ion_prior_train_fdr: TDC q-value threshold of the training PSMs
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add c and z+1 ions for electron-activated spectra
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report:isotope_error_convention: the PSMs report isotope_error as observed minus theoretical (true) or with the sign of the search offset, theoretical minus observed (false)
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mutableprotected |
Most recent calibration result (valid after any search that invoked runCalibrationPass_). Stored for test observability and diagnostics. Marked mutable because it is pure diagnostic/telemetry state that doesn't affect the logical const-ness of search().
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mutableprotected |
Scalar tolerance passed to OpenSearchModificationAnalysis on the most recent search() call. Stored for test observability: because the calibration writeback restores the tolerance members on exit (to avoid per-file state leaks in the multi-file wrapper), tests that want to verify "the mod analyzer received the calibrated value, not the user-configured one" can't just read the members post-search — they need to see what was actually passed to the analyzer. Default -1.0 (sentinel: no search has run yet).
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mutableprotected |
Percolator rescoring outcome of the most recent search(spectra, ctx, ...) call, bridged to SearchResult::percolator as last_run_stats_ is to SearchResult::stats.
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mutableprotected |
Per-run statistics of the most recent search(spectra, ctx, ...) call. Bridges the const search() (which returns ExitCodes, not a SearchResult) to its callers, which copy this into SearchResult::stats. Reset at the start of each search() call. mutable for the same reason as above: pure diagnostic state, orthogonal to logical const-ness.
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Resolved scoring:method: mass-accuracy weighted HyperScore.
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Width of the mass-accuracy kernel in ppm (scoring:mass_error_sd)
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staticconstexpr |
See MIN_DECOYS_PER_TARGET.
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percolator:max_train_psms (0: no cap)
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staticconstexpr |
With 'decoys' auto, the decoys of the database are reused only if there are at least this many decoys per target protein (and at most MAX_DECOYS_PER_TARGET); otherwise the search refuses the database (Exception::InvalidParameter)
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mutableprotected |
both switched off by buildDecoyAugmentedDB_()
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NLargest cap on MS2 peaks before scoring; 0 = resolution-aware auto (peaks:keep_n)
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WindowMower peaks-per-100Da before scoring (peaks:window_top)
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Resolution-aware treatment of the final peak window.
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rescore_method 'reset': Percolator's RESET parameters (empty: Percolator)
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rescoring:...: options of Percolator's training
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mutableprotected |
With pool_rescoring_: the input files of the multi-file search (for the share of the transfer model's training PSMs that a file rescored on its own keeps, transferQuota_())
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mutableprotected |
Set by the multi-file searches with 'rescore' while their files are searched: the PSMs of each file before the rescoring are kept (SearchResult::pre_fdr_peptide_ids) for rescoreFilesTogether_()
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mutableprotected |
The PSMs of the most recent search(spectra, ctx, ...) before FDR:PSM (only with keep_pre_fdr_psms_), bridged to the caller like last_run_stats_.
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mutableprotected |
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mutableprotected |
positive magnitude (default matches the param)
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peptide:protein_mapping = index
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Retrieve candidates with the spectrum before deisotoping and local filtering (fragment:query_spectrum)
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mutableprotected |
The hits that the most recent rescoring dropped (all but the best per spectrum), destroyed by a helper thread meanwhile (see applyPsmFDR_()); the holder joins it when it goes (at the next rescoring of this object, or with the last copy of this object that shares it).
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rescore: Percolator rescoring of every run before its PSM-level FDR
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rescoring:top_hits: the hits per spectrum that the Percolator rescoring gets (0 = report_top_hits_)
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mutableprotected |
spectra file of the run being rescored (names its RESET stem table)
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mutableprotected |
rescore_reuse: the current search defers its rescoring and FDR:PSM to finishDeferredRescoring_()
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mutableprotected |
rescore_reuse: the index of the file of the current search in a multi-file search (0 otherwise)
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protected |
rescore_reuse:max_shift
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rescore_reuse:max_train
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protected |
rescore_reuse:mode
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mutableprotected |
rescore_reuse: the models of a multi-file search ('static': trained on its first files; null otherwise)
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protected |
rescore_reuse:static_files
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protected |
Upper fragment charge bound (fragment:max_charge)
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protected |
Resolved scoring:fragment_charges: score charges above 1.
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staticconstexprprotected |
ProSE's own PSM score (HyperScore, in log space): the main score of its PSMs, and after Percolator rescoring or FDR:PSM a meta value of this name (or with the suffix "_score", as FalseDiscoveryRate keeps it)
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protected |
annotate:self_trained_ion_priors
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mutableprotected |
rescoring:stratum_q_values (effective)
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mutableprotected |
rescoring:terminal_features (effective)
1.9.8