// *************************************************************************** // BamMultiReader.cpp (c) 2010 Erik Garrison, Derek Barnett // Marth Lab, Department of Biology, Boston College // All rights reserved. // --------------------------------------------------------------------------- // Last modified: 18 September 2010 (DB) // --------------------------------------------------------------------------- // Uses BGZF routines were adapted from the bgzf.c code developed at the Broad // Institute. // --------------------------------------------------------------------------- // Functionality for simultaneously reading multiple BAM files. // // This functionality allows applications to work on very large sets of files // without requiring intermediate merge, sort, and index steps for each file // subset. It also improves the performance of our merge system as it // precludes the need to sort merged files. // *************************************************************************** #include #include #include #include #include #include #include #include "BGZF.h" #include "BamMultiReader.h" using namespace BamTools; using namespace std; // ----------------------------------------------------- // BamMultiReader implementation // ----------------------------------------------------- // constructor BamMultiReader::BamMultiReader(void) : CurrentRefID(0) , CurrentLeft(0) { } // destructor BamMultiReader::~BamMultiReader(void) { Close(); } // close the BAM files void BamMultiReader::Close(void) { // close all BAM readers and clean up pointers vector >::iterator readerIter = readers.begin(); vector >::iterator readerEnd = readers.end(); for ( ; readerIter != readerEnd; ++readerIter) { BamReader* reader = (*readerIter).first; BamAlignment* alignment = (*readerIter).second; // close the reader if ( reader) reader->Close(); // delete reader pointer delete reader; reader = 0; // delete alignment pointer delete alignment; alignment = 0; } // clear out the container readers.clear(); } // saves index data to BAM index files (".bai"/".bti") where necessary, returns success/fail bool BamMultiReader::CreateIndexes(bool useStandardIndex) { bool result = true; for (vector >::iterator it = readers.begin(); it != readers.end(); ++it) { BamReader* reader = it->first; result &= reader->CreateIndex(useStandardIndex); } return result; } // for debugging void BamMultiReader::DumpAlignmentIndex(void) { for (AlignmentIndex::const_iterator it = alignments.begin(); it != alignments.end(); ++it) { cerr << it->first.first << ":" << it->first.second << " " << it->second.first->GetFilename() << endl; } } // makes a virtual, unified header for all the bam files in the multireader const string BamMultiReader::GetHeaderText(void) const { string mergedHeader = ""; map readGroups; // foreach extraction entry (each BAM file) for (vector >::const_iterator rs = readers.begin(); rs != readers.end(); ++rs) { BamReader* reader = rs->first; const string headerText = reader->GetHeaderText(); if ( headerText.empty() ) continue; map currentFileReadGroups; stringstream header(headerText); vector lines; string item; while (getline(header, item)) lines.push_back(item); for (vector::const_iterator it = lines.begin(); it != lines.end(); ++it) { // get next line from header, skip if empty string headerLine = *it; if ( headerLine.empty() ) { continue; } // if first file, save HD & SQ entries if ( rs == readers.begin() ) { if ( headerLine.find("@HD") == 0 || headerLine.find("@SQ") == 0) { mergedHeader.append(headerLine.c_str()); mergedHeader.append(1, '\n'); } } // (for all files) append RG entries if they are unique if ( headerLine.find("@RG") == 0 ) { stringstream headerLineSs(headerLine); string part, readGroupPart, readGroup; while(std::getline(headerLineSs, part, '\t')) { stringstream partSs(part); string subtag; std::getline(partSs, subtag, ':'); if (subtag == "ID") { std::getline(partSs, readGroup, ':'); break; } } if (readGroups.find(readGroup) == readGroups.end()) { // prevents duplicate @RG entries mergedHeader.append(headerLine.c_str() ); mergedHeader.append(1, '\n'); readGroups[readGroup] = true; currentFileReadGroups[readGroup] = true; } else { // warn iff we are reading one file and discover duplicated @RG tags in the header // otherwise, we emit no warning, as we might be merging multiple BAM files with identical @RG tags if (currentFileReadGroups.find(readGroup) != currentFileReadGroups.end()) { cerr << "WARNING: duplicate @RG tag " << readGroup << " entry in header of " << reader->GetFilename() << endl; } } } } } // return merged header text return mergedHeader; } // get next alignment among all files bool BamMultiReader::GetNextAlignment(BamAlignment& nextAlignment) { // bail out if we are at EOF in all files, means no more alignments to process if (!HasOpenReaders()) return false; // when all alignments have stepped into a new target sequence, update our // current reference sequence id UpdateReferenceID(); // our lowest alignment and reader will be at the front of our alignment index BamAlignment* alignment = alignments.begin()->second.second; BamReader* reader = alignments.begin()->second.first; // now that we have the lowest alignment in the set, save it by copy to our argument nextAlignment = BamAlignment(*alignment); // remove this alignment index entry from our alignment index alignments.erase(alignments.begin()); // and add another entry if we can get another alignment from the reader if (reader->GetNextAlignment(*alignment)) { alignments.insert(make_pair(make_pair(alignment->RefID, alignment->Position), make_pair(reader, alignment))); } else { // do nothing //cerr << "reached end of file " << lowestReader->GetFilename() << endl; } return true; } // get next alignment among all files without parsing character data from alignments bool BamMultiReader::GetNextAlignmentCore(BamAlignment& nextAlignment) { // bail out if we are at EOF in all files, means no more alignments to process if (!HasOpenReaders()) return false; // when all alignments have stepped into a new target sequence, update our // current reference sequence id UpdateReferenceID(); // our lowest alignment and reader will be at the front of our alignment index BamAlignment* alignment = alignments.begin()->second.second; BamReader* reader = alignments.begin()->second.first; // now that we have the lowest alignment in the set, save it by copy to our argument nextAlignment = BamAlignment(*alignment); //memcpy(&nextAlignment, alignment, sizeof(BamAlignment)); // remove this alignment index entry from our alignment index alignments.erase(alignments.begin()); // and add another entry if we can get another alignment from the reader if (reader->GetNextAlignmentCore(*alignment)) { alignments.insert(make_pair(make_pair(alignment->RefID, alignment->Position), make_pair(reader, alignment))); } else { // do nothing //cerr << "reached end of file " << lowestReader->GetFilename() << endl; } return true; } // --------------------------------------------------------------------------------------- // // NB: The following GetReferenceX() functions assume that we have identical // references for all BAM files. We enforce this by invoking the above // validation function (ValidateReaders) to verify that our reference data // is the same across all files on Open, so we will not encounter a situation // in which there is a mismatch and we are still live. // // --------------------------------------------------------------------------------------- // returns the number of reference sequences const int BamMultiReader::GetReferenceCount(void) const { return readers.front().first->GetReferenceCount(); } // returns vector of reference objects const BamTools::RefVector BamMultiReader::GetReferenceData(void) const { return readers.front().first->GetReferenceData(); } // returns refID from reference name const int BamMultiReader::GetReferenceID(const string& refName) const { return readers.front().first->GetReferenceID(refName); } // --------------------------------------------------------------------------------------- // checks if any readers still have alignments bool BamMultiReader::HasOpenReaders() { return alignments.size() > 0; } // returns whether underlying BAM readers ALL have an index loaded // this is useful to indicate whether Jump() or SetRegion() are possible bool BamMultiReader::IsIndexLoaded(void) const { bool ok = true; vector >::const_iterator readerIter = readers.begin(); vector >::const_iterator readerEnd = readers.end(); for ( ; readerIter != readerEnd; ++readerIter ) { const BamReader* reader = (*readerIter).first; if ( reader ) ok &= reader->IsIndexLoaded(); } return ok; } // jumps to specified region(refID, leftBound) in BAM files, returns success/fail bool BamMultiReader::Jump(int refID, int position) { //if ( References.at(refID).RefHasAlignments && (position <= References.at(refID).RefLength) ) { CurrentRefID = refID; CurrentLeft = position; bool result = true; for (vector >::iterator it = readers.begin(); it != readers.end(); ++it) { BamReader* reader = it->first; result &= reader->Jump(refID, position); if (!result) { cerr << "ERROR: could not jump " << reader->GetFilename() << " to " << refID << ":" << position << endl; exit(1); } } if (result) UpdateAlignments(); return result; } // opens BAM files bool BamMultiReader::Open(const vector& filenames, bool openIndexes, bool coreMode, bool useDefaultIndex) { // for filename in filenames fileNames = filenames; // save filenames in our multireader for (vector::const_iterator it = filenames.begin(); it != filenames.end(); ++it) { const string filename = *it; BamReader* reader = new BamReader; bool openedOK = true; if (openIndexes) { // leave index filename empty // this allows BamReader & BamIndex to search for any available // useDefaultIndex gives hint to prefer BAI over BTI openedOK = reader->Open(filename, "", true, useDefaultIndex); } // ignoring index file(s) else openedOK = reader->Open(filename); // if file opened ok, check that it can be read if ( openedOK ) { bool fileOK = true; BamAlignment* alignment = new BamAlignment; fileOK &= ( coreMode ? reader->GetNextAlignmentCore(*alignment) : reader->GetNextAlignment(*alignment) ); if (fileOK) { readers.push_back(make_pair(reader, alignment)); // store pointers to our readers for cleanup alignments.insert(make_pair(make_pair(alignment->RefID, alignment->Position), make_pair(reader, alignment))); } else { cerr << "WARNING: could not read first alignment in " << filename << ", ignoring file" << endl; // if only file available & could not be read, return failure if ( filenames.size() == 1 ) return false; } } // TODO; any further error handling when openedOK is false ?? else return false; } // files opened ok, at least one alignment could be read, // now need to check that all files use same reference data ValidateReaders(); return true; } void BamMultiReader::PrintFilenames(void) { for (vector >::iterator it = readers.begin(); it != readers.end(); ++it) { BamReader* reader = it->first; cout << reader->GetFilename() << endl; } } // returns BAM file pointers to beginning of alignment data bool BamMultiReader::Rewind(void) { bool result = true; for (vector >::iterator it = readers.begin(); it != readers.end(); ++it) { BamReader* reader = it->first; result &= reader->Rewind(); } return result; } bool BamMultiReader::SetRegion(const int& leftRefID, const int& leftPosition, const int& rightRefID, const int& rightPosition) { BamRegion region(leftRefID, leftPosition, rightRefID, rightPosition); return SetRegion(region); } bool BamMultiReader::SetRegion(const BamRegion& region) { Region = region; // NB: While it may make sense to track readers in which we can // successfully SetRegion, In practice a failure of SetRegion means "no // alignments here." It makes sense to simply accept the failure, // UpdateAlignments(), and continue. for (vector >::iterator it = readers.begin(); it != readers.end(); ++it) { if (!it->first->SetRegion(region)) { cerr << "ERROR: could not jump " << it->first->GetFilename() << " to " << region.LeftRefID << ":" << region.LeftPosition << ".." << region.RightRefID << ":" << region.RightPosition << endl; } } UpdateAlignments(); return true; } void BamMultiReader::UpdateAlignments(void) { // Update Alignments alignments.clear(); for (vector >::iterator it = readers.begin(); it != readers.end(); ++it) { BamReader* br = it->first; BamAlignment* ba = it->second; if (br->GetNextAlignment(*ba)) { alignments.insert(make_pair(make_pair(ba->RefID, ba->Position), make_pair(br, ba))); } else { // assume BamReader end of region / EOF } } } // updates the reference id stored in the BamMultiReader // to reflect the current state of the readers void BamMultiReader::UpdateReferenceID(void) { // the alignments are sorted by position, so the first alignment will always have the lowest reference ID if (alignments.begin()->second.second->RefID != CurrentRefID) { // get the next reference id // while there aren't any readers at the next ref id // increment the ref id int nextRefID = CurrentRefID; while (alignments.begin()->second.second->RefID != nextRefID) { ++nextRefID; } //cerr << "updating reference id from " << CurrentRefID << " to " << nextRefID << endl; CurrentRefID = nextRefID; } } // ValidateReaders checks that all the readers point to BAM files representing // alignments against the same set of reference sequences, and that the // sequences are identically ordered. If these checks fail the operation of // the multireader is undefined, so we force program exit. void BamMultiReader::ValidateReaders(void) const { int firstRefCount = readers.front().first->GetReferenceCount(); BamTools::RefVector firstRefData = readers.front().first->GetReferenceData(); for (vector >::const_iterator it = readers.begin(); it != readers.end(); ++it) { BamReader* reader = it->first; BamTools::RefVector currentRefData = reader->GetReferenceData(); BamTools::RefVector::const_iterator f = firstRefData.begin(); BamTools::RefVector::const_iterator c = currentRefData.begin(); if (reader->GetReferenceCount() != firstRefCount || firstRefData.size() != currentRefData.size()) { cerr << "ERROR: mismatched number of references in " << reader->GetFilename() << " expected " << firstRefCount << " reference sequences but only found " << reader->GetReferenceCount() << endl; exit(1); } // this will be ok; we just checked above that we have identically-sized sets of references // here we simply check if they are all, in fact, equal in content while (f != firstRefData.end()) { if (f->RefName != c->RefName || f->RefLength != c->RefLength) { cerr << "ERROR: mismatched references found in " << reader->GetFilename() << " expected: " << endl; for (BamTools::RefVector::const_iterator a = firstRefData.begin(); a != firstRefData.end(); ++a) cerr << a->RefName << " " << a->RefLength << endl; cerr << "but found: " << endl; for (BamTools::RefVector::const_iterator a = currentRefData.begin(); a != currentRefData.end(); ++a) cerr << a->RefName << " " << a->RefLength << endl; exit(1); } ++f; ++c; } } }