1 // ***************************************************************************
2 // bamtools_convert.cpp (c) 2010 Derek Barnett, Erik Garrison
3 // Marth Lab, Department of Biology, Boston College
4 // All rights reserved.
5 // ---------------------------------------------------------------------------
6 // Last modified: 23 September 2010
7 // ---------------------------------------------------------------------------
8 // Converts between BAM and a number of other formats
9 // ***************************************************************************
16 #include "bamtools_convert.h"
17 #include "bamtools_fasta.h"
18 #include "bamtools_options.h"
19 #include "bamtools_pileup_engine.h"
20 #include "bamtools_utilities.h"
22 #include "BamMultiReader.h"
24 using namespace BamTools;
28 // ---------------------------------------------
29 // ConvertTool constants
31 // supported conversion format command-line names
32 static const string FORMAT_BED = "bed";
33 static const string FORMAT_BEDGRAPH = "bedgraph";
34 static const string FORMAT_FASTA = "fasta";
35 static const string FORMAT_FASTQ = "fastq";
36 static const string FORMAT_JSON = "json";
37 static const string FORMAT_SAM = "sam";
38 static const string FORMAT_PILEUP = "pileup";
39 static const string FORMAT_WIGGLE = "wig";
42 static const unsigned int FASTA_LINE_MAX = 50;
44 // ---------------------------------------------
45 // ConvertPileupFormatVisitor declaration
47 class ConvertPileupFormatVisitor : public PileupVisitor {
51 ConvertPileupFormatVisitor(const RefVector& references,
52 const string& fastaFilename,
53 const bool isPrintingMapQualities,
55 ~ConvertPileupFormatVisitor(void);
57 // PileupVisitor interface implementation
59 void Visit(const PileupPosition& pileupData);
65 bool m_isPrintingMapQualities;
67 RefVector m_references;
70 } // namespace BamTools
72 // ---------------------------------------------
73 // ConvertSettings implementation
75 struct ConvertTool::ConvertSettings {
84 bool HasFastaFilename;
85 bool IsOmittingSamHeader;
86 bool IsPrintingPileupMapQualities;
89 vector<string> InputFiles;
90 string OutputFilename;
103 , HasFastaFilename(false)
104 , IsOmittingSamHeader(false)
105 , IsPrintingPileupMapQualities(false)
106 , OutputFilename(Options::StandardOut())
111 // ---------------------------------------------
112 // ConvertToolPrivate implementation
114 struct ConvertTool::ConvertToolPrivate {
118 ConvertToolPrivate(ConvertTool::ConvertSettings* settings);
119 ~ConvertToolPrivate(void);
127 void PrintBed(const BamAlignment& a);
128 void PrintBedGraph(const BamAlignment& a);
129 void PrintFasta(const BamAlignment& a);
130 void PrintFastq(const BamAlignment& a);
131 void PrintJson(const BamAlignment& a);
132 void PrintSam(const BamAlignment& a);
133 void PrintWiggle(const BamAlignment& a);
135 // special case - uses the PileupEngine
136 bool RunPileupConversion(BamMultiReader* reader);
140 ConvertTool::ConvertSettings* m_settings;
141 RefVector m_references;
145 ConvertTool::ConvertToolPrivate::ConvertToolPrivate(ConvertTool::ConvertSettings* settings)
146 : m_settings(settings)
147 , m_out(cout.rdbuf()) // default output to cout
150 ConvertTool::ConvertToolPrivate::~ConvertToolPrivate(void) { }
152 bool ConvertTool::ConvertToolPrivate::Run(void) {
154 // ------------------------------------
155 // initialize conversion input/output
157 // set to default input if none provided
158 if ( !m_settings->HasInput )
159 m_settings->InputFiles.push_back(Options::StandardIn());
162 BamMultiReader reader;
163 reader.Open(m_settings->InputFiles);
164 m_references = reader.GetReferenceData();
166 // set region if specified
168 if ( m_settings->HasRegion ) {
169 if ( Utilities::ParseRegionString(m_settings->Region, reader, region) ) {
170 if ( !reader.SetRegion(region) ) {
171 cerr << "Could not set BamReader region to REGION: " << m_settings->Region << endl;
175 cerr << "Could not parse REGION: " << m_settings->Region << endl;
180 // if output file given
182 if ( m_settings->HasOutput ) {
184 // open output file stream
185 outFile.open(m_settings->OutputFilename.c_str());
187 cerr << "Could not open " << m_settings->OutputFilename << " for output" << endl;
191 // set m_out to file's streambuf
192 m_out.rdbuf(outFile.rdbuf());
195 // -------------------------------------
196 // do conversion based on format
198 bool convertedOk = true;
200 // pileup is special case
201 // conversion not done per alignment, like the other formats
202 if ( m_settings->Format == FORMAT_PILEUP )
203 convertedOk = RunPileupConversion(&reader);
208 bool formatError = false;
210 // set function pointer to proper conversion method
211 void (BamTools::ConvertTool::ConvertToolPrivate::*pFunction)(const BamAlignment&) = 0;
212 if ( m_settings->Format == FORMAT_BED ) pFunction = &BamTools::ConvertTool::ConvertToolPrivate::PrintBed;
213 else if ( m_settings->Format == FORMAT_BEDGRAPH ) pFunction = &BamTools::ConvertTool::ConvertToolPrivate::PrintBedGraph;
214 else if ( m_settings->Format == FORMAT_FASTA ) pFunction = &BamTools::ConvertTool::ConvertToolPrivate::PrintFasta;
215 else if ( m_settings->Format == FORMAT_FASTQ ) pFunction = &BamTools::ConvertTool::ConvertToolPrivate::PrintFastq;
216 else if ( m_settings->Format == FORMAT_JSON ) pFunction = &BamTools::ConvertTool::ConvertToolPrivate::PrintJson;
217 else if ( m_settings->Format == FORMAT_SAM ) pFunction = &BamTools::ConvertTool::ConvertToolPrivate::PrintSam;
218 else if ( m_settings->Format == FORMAT_WIGGLE ) pFunction = &BamTools::ConvertTool::ConvertToolPrivate::PrintWiggle;
220 cerr << "Unrecognized format: " << m_settings->Format << endl;
221 cerr << "Please see help|README (?) for details on supported formats " << endl;
226 // if format selected ok
227 if ( !formatError ) {
229 // if SAM format & not omitting header, print SAM header first
230 if ( (m_settings->Format == FORMAT_SAM) && !m_settings->IsOmittingSamHeader )
231 m_out << reader.GetHeaderText();
233 // iterate through file, doing conversion
235 while ( reader.GetNextAlignment(a) )
236 (this->*pFunction)(a);
238 // set flag for successful conversion
243 // ------------------------
247 if ( m_settings->HasOutput ) outFile.close();
251 // ----------------------------------------------------------
252 // Conversion/output methods
253 // ----------------------------------------------------------
255 void ConvertTool::ConvertToolPrivate::PrintBed(const BamAlignment& a) {
257 // tab-delimited, 0-based half-open
258 // (e.g. a 50-base read aligned to pos 10 could have BED coordinates (10, 60) instead of BAM coordinates (10, 59) )
259 // <chromName> <chromStart> <chromEnd> <readName> <score> <strand>
261 m_out << m_references.at(a.RefID).RefName << "\t"
262 << a.Position << "\t"
263 << a.GetEndPosition() + 1 << "\t"
265 << a.MapQuality << "\t"
266 << (a.IsReverseStrand() ? "-" : "+") << endl;
269 void ConvertTool::ConvertToolPrivate::PrintBedGraph(const BamAlignment& a) {
273 // print BamAlignment in FASTA format
274 // N.B. - uses QueryBases NOT AlignedBases
275 void ConvertTool::ConvertToolPrivate::PrintFasta(const BamAlignment& a) {
277 // >BamAlignment.Name
278 // BamAlignment.QueryBases (up to FASTA_LINE_MAX bases per line)
281 // N.B. - QueryBases are reverse-complemented if aligned to reverse strand
284 m_out << "> " << a.Name << endl;
286 // handle reverse strand alignment - bases
287 string sequence = a.QueryBases;
288 if ( a.IsReverseStrand() )
289 Utilities::ReverseComplement(sequence);
291 // if sequence fits on single line
292 if ( sequence.length() <= FASTA_LINE_MAX )
293 m_out << sequence << endl;
295 // else split over multiple lines
299 size_t seqLength = sequence.length(); // handle reverse strand alignment - bases & qualitiesth();
301 // write subsequences to each line
302 while ( position < (seqLength - FASTA_LINE_MAX) ) {
303 m_out << sequence.substr(position, FASTA_LINE_MAX) << endl;
304 position += FASTA_LINE_MAX;
307 // write final subsequence
308 m_out << sequence.substr(position) << endl;
312 // print BamAlignment in FASTQ format
313 // N.B. - uses QueryBases NOT AlignedBases
314 void ConvertTool::ConvertToolPrivate::PrintFastq(const BamAlignment& a) {
316 // @BamAlignment.Name
317 // BamAlignment.QueryBases
319 // BamAlignment.Qualities
321 // N.B. - QueryBases are reverse-complemented (& Qualities reversed) if aligned to reverse strand .
322 // Name is appended "/1" or "/2" if paired-end, to reflect which mate this entry is.
324 // handle paired-end alignments
325 string name = a.Name;
327 name.append( (a.IsFirstMate() ? "/1" : "/2") );
329 // handle reverse strand alignment - bases & qualities
330 string qualities = a.Qualities;
331 string sequence = a.QueryBases;
332 if ( a.IsReverseStrand() ) {
333 Utilities::Reverse(qualities);
334 Utilities::ReverseComplement(sequence);
337 // write to output stream
338 m_out << "@" << name << endl
341 << qualities << endl;
344 // print BamAlignment in JSON format
345 void ConvertTool::ConvertToolPrivate::PrintJson(const BamAlignment& a) {
347 // write name & alignment flag
348 m_out << "{\"name\":\"" << a.Name << "\",\"alignmentFlag\":\"" << a.AlignmentFlag << "\",";
350 // write reference name
351 if ( (a.RefID >= 0) && (a.RefID < (int)m_references.size()) )
352 m_out << "\"reference\":\"" << m_references[a.RefID].RefName << "\",";
354 // write position & map quality
355 m_out << "\"position\":" << a.Position+1 << ",\"mapQuality\":" << a.MapQuality << ",";
358 const vector<CigarOp>& cigarData = a.CigarData;
359 if ( !cigarData.empty() ) {
360 m_out << "\"cigar\":[";
361 vector<CigarOp>::const_iterator cigarBegin = cigarData.begin();
362 vector<CigarOp>::const_iterator cigarIter = cigarBegin;
363 vector<CigarOp>::const_iterator cigarEnd = cigarData.end();
364 for ( ; cigarIter != cigarEnd; ++cigarIter ) {
365 const CigarOp& op = (*cigarIter);
366 if (cigarIter != cigarBegin) m_out << ",";
367 m_out << "\"" << op.Length << op.Type << "\"";
372 // write mate reference name, mate position, & insert size
373 if ( a.IsPaired() && (a.MateRefID >= 0) && (a.MateRefID < (int)m_references.size()) ) {
374 m_out << "\"mate\":{"
375 << "\"reference\":\"" << m_references[a.MateRefID].RefName << "\","
376 << "\"position\":" << a.MatePosition+1
377 << ",\"insertSize\":" << a.InsertSize << "},";
381 if ( !a.QueryBases.empty() )
382 m_out << "\"queryBases\":\"" << a.QueryBases << "\",";
385 if ( !a.Qualities.empty() ) {
386 string::const_iterator s = a.Qualities.begin();
387 m_out << "\"qualities\":[" << static_cast<short>(*s) - 33;
389 for (; s != a.Qualities.end(); ++s) {
390 m_out << "," << static_cast<short>(*s) - 33;
396 const char* tagData = a.TagData.c_str();
397 const size_t tagDataLength = a.TagData.length();
399 if (index < tagDataLength) {
401 m_out << "\"tags\":{";
403 while ( index < tagDataLength ) {
409 m_out << "\"" << a.TagData.substr(index, 2) << "\":";
413 char type = a.TagData.at(index);
418 m_out << "\"" << tagData[index] << "\"";
423 m_out << (int)tagData[index];
428 m_out << (int)tagData[index];
433 m_out << BgzfData::UnpackUnsignedShort(&tagData[index]);
438 m_out << BgzfData::UnpackSignedShort(&tagData[index]);
443 m_out << BgzfData::UnpackUnsignedInt(&tagData[index]);
448 m_out << BgzfData::UnpackSignedInt(&tagData[index]);
453 m_out << BgzfData::UnpackFloat(&tagData[index]);
458 m_out << BgzfData::UnpackDouble(&tagData[index]);
465 while (tagData[index]) {
466 m_out << tagData[index];
474 if ( tagData[index] == '\0')
481 m_out << "}" << endl;
484 // print BamAlignment in SAM format
485 void ConvertTool::ConvertToolPrivate::PrintSam(const BamAlignment& a) {
488 // <QNAME> <FLAG> <RNAME> <POS> <MAPQ> <CIGAR> <MRNM> <MPOS> <ISIZE> <SEQ> <QUAL> [ <TAG>:<VTYPE>:<VALUE> [...] ]
490 // write name & alignment flag
491 m_out << a.Name << "\t" << a.AlignmentFlag << "\t";
493 // write reference name
494 if ( (a.RefID >= 0) && (a.RefID < (int)m_references.size()) )
495 m_out << m_references[a.RefID].RefName << "\t";
499 // write position & map quality
500 m_out << a.Position+1 << "\t" << a.MapQuality << "\t";
503 const vector<CigarOp>& cigarData = a.CigarData;
504 if ( cigarData.empty() ) m_out << "*\t";
506 vector<CigarOp>::const_iterator cigarIter = cigarData.begin();
507 vector<CigarOp>::const_iterator cigarEnd = cigarData.end();
508 for ( ; cigarIter != cigarEnd; ++cigarIter ) {
509 const CigarOp& op = (*cigarIter);
510 m_out << op.Length << op.Type;
515 // write mate reference name, mate position, & insert size
516 if ( a.IsPaired() && (a.MateRefID >= 0) && (a.MateRefID < (int)m_references.size()) ) {
517 if ( a.MateRefID == a.RefID ) m_out << "=\t";
518 else m_out << m_references[a.MateRefID].RefName << "\t";
519 m_out << a.MatePosition+1 << "\t" << a.InsertSize << "\t";
521 else m_out << "*\t0\t0\t";
524 if ( a.QueryBases.empty() ) m_out << "*\t";
525 else m_out << a.QueryBases << "\t";
528 if ( a.Qualities.empty() ) m_out << "*";
529 else m_out << a.Qualities;
532 const char* tagData = a.TagData.c_str();
533 const size_t tagDataLength = a.TagData.length();
536 while ( index < tagDataLength ) {
539 string tagName = a.TagData.substr(index, 2);
540 m_out << "\t" << tagName << ":";
544 char type = a.TagData.at(index);
548 m_out << "A:" << tagData[index];
553 m_out << "i:" << (int)tagData[index];
558 m_out << "i:" << (int)tagData[index];
563 m_out << "i:" << BgzfData::UnpackUnsignedShort(&tagData[index]);
568 m_out << "i:" << BgzfData::UnpackSignedShort(&tagData[index]);
573 m_out << "i:" << BgzfData::UnpackUnsignedInt(&tagData[index]);
578 m_out << "i:" << BgzfData::UnpackSignedInt(&tagData[index]);
583 m_out << "f:" << BgzfData::UnpackFloat(&tagData[index]);
588 m_out << "d:" << BgzfData::UnpackDouble(&tagData[index]);
594 m_out << type << ":";
595 while (tagData[index]) {
596 m_out << tagData[index];
603 if ( tagData[index] == '\0')
610 void ConvertTool::ConvertToolPrivate::PrintWiggle(const BamAlignment& a) {
614 bool ConvertTool::ConvertToolPrivate::RunPileupConversion(BamMultiReader* reader) {
616 // check for valid BamMultiReader
617 if ( reader == 0 ) return false;
619 // set up our pileup format 'visitor'
620 ConvertPileupFormatVisitor* v = new ConvertPileupFormatVisitor(m_references,
621 m_settings->FastaFilename,
622 m_settings->IsPrintingPileupMapQualities,
625 // set up PileupEngine
627 pileup.AddVisitor(v);
629 // iterate through data
631 while ( reader->GetNextAlignment(al) )
632 pileup.AddAlignment(al);
643 // ---------------------------------------------
644 // ConvertTool implementation
646 ConvertTool::ConvertTool(void)
648 , m_settings(new ConvertSettings)
651 // set program details
652 Options::SetProgramInfo("bamtools convert", "converts BAM to a number of other formats", "-format <FORMAT> [-in <filename> -in <filename> ...] [-out <filename>] [other options]");
655 OptionGroup* IO_Opts = Options::CreateOptionGroup("Input & Output");
656 Options::AddValueOption("-in", "BAM filename", "the input BAM file(s)", "", m_settings->HasInput, m_settings->InputFiles, IO_Opts, Options::StandardIn());
657 Options::AddValueOption("-out", "BAM filename", "the output BAM file", "", m_settings->HasOutput, m_settings->OutputFilename, IO_Opts, Options::StandardOut());
658 Options::AddValueOption("-format", "FORMAT", "the output file format - see README for recognized formats", "", m_settings->HasFormat, m_settings->Format, IO_Opts);
660 OptionGroup* FilterOpts = Options::CreateOptionGroup("Filters");
661 Options::AddValueOption("-region", "REGION", "genomic region. Index file is recommended for better performance, and is read automatically if it exists as <filename>.bai. See \'bamtools help index\' for more details on creating one", "", m_settings->HasRegion, m_settings->Region, FilterOpts);
663 OptionGroup* PileupOpts = Options::CreateOptionGroup("Pileup Options");
664 Options::AddValueOption("-fasta", "FASTA filename", "FASTA reference file", "", m_settings->HasFastaFilename, m_settings->FastaFilename, PileupOpts, "");
665 Options::AddOption("-mapqual", "print the mapping qualities", m_settings->IsPrintingPileupMapQualities, PileupOpts);
667 OptionGroup* SamOpts = Options::CreateOptionGroup("SAM Options");
668 Options::AddOption("-noheader", "omit the SAM header from output", m_settings->IsOmittingSamHeader, SamOpts);
671 ConvertTool::~ConvertTool(void) {
679 int ConvertTool::Help(void) {
680 Options::DisplayHelp();
684 int ConvertTool::Run(int argc, char* argv[]) {
686 // parse command line arguments
687 Options::Parse(argc, argv, 1);
689 // run internal ConvertTool implementation, return success/fail
690 m_impl = new ConvertToolPrivate(m_settings);
698 // ---------------------------------------------
699 // ConvertPileupFormatVisitor implementation
701 ConvertPileupFormatVisitor::ConvertPileupFormatVisitor(const RefVector& references,
702 const string& fastaFilename,
703 const bool isPrintingMapQualities,
707 , m_isPrintingMapQualities(isPrintingMapQualities)
709 , m_references(references)
711 // set up Fasta reader if file is provided
712 if ( !fastaFilename.empty() ) {
714 // check for FASTA index
715 string indexFilename = "";
716 if ( Utilities::FileExists(fastaFilename + ".fai") )
717 indexFilename = fastaFilename + ".fai";
720 if ( m_fasta.Open(fastaFilename, indexFilename) )
725 ConvertPileupFormatVisitor::~ConvertPileupFormatVisitor(void) {
726 // be sure to close Fasta reader
733 void ConvertPileupFormatVisitor::Visit(const PileupPosition& pileupData ) {
735 // skip if no alignments at this position
736 if ( pileupData.PileupAlignments.empty() ) return;
738 // retrieve reference name
739 const string& referenceName = m_references[pileupData.RefId].RefName;
740 const int& position = pileupData.Position;
742 // retrieve reference base from FASTA file, if one provided; otherwise default to 'N'
743 char referenceBase('N');
744 if ( m_hasFasta && (pileupData.Position < m_references[pileupData.RefId].RefLength) ) {
745 if ( !m_fasta.GetBase(pileupData.RefId, pileupData.Position, referenceBase ) ) {
746 cerr << "Pileup error : Could not read reference base from FASTA file" << endl;
751 // get count of alleles at this position
752 const int numberAlleles = pileupData.PileupAlignments.size();
754 // -----------------------------------------------------------
755 // build strings based on alleles at this positionInAlignment
757 stringstream bases("");
758 stringstream baseQualities("");
759 stringstream mapQualities("");
761 // iterate over alignments at this pileup position
762 vector<PileupAlignment>::const_iterator pileupIter = pileupData.PileupAlignments.begin();
763 vector<PileupAlignment>::const_iterator pileupEnd = pileupData.PileupAlignments.end();
764 for ( ; pileupIter != pileupEnd; ++pileupIter ) {
765 const PileupAlignment pa = (*pileupIter);
766 const BamAlignment& ba = pa.Alignment;
768 // if beginning of read segment
769 if ( pa.IsSegmentBegin )
770 bases << '^' << ( ((int)ba.MapQuality > 93) ? (char)126 : (char)((int)ba.MapQuality+33) );
772 // if current base is not a DELETION
773 if ( !pa.IsCurrentDeletion ) {
775 // get base at current position
776 char base = ba.QueryBases.at(pa.PositionInAlignment);
778 // if base matches reference
780 toupper(base) == toupper(referenceBase) ||
781 tolower(base) == tolower(referenceBase) )
783 base = (ba.IsReverseStrand() ? ',' : '.' );
786 // mismatches reference
787 else base = (ba.IsReverseStrand() ? tolower(base) : toupper(base) );
792 // if next position contains insertion
793 if ( pa.IsNextInsertion ) {
794 bases << '+' << pa.InsertionLength;
795 for (int i = 1; i <= pa.InsertionLength; ++i) {
796 char insertedBase = (char)ba.QueryBases.at(pa.PositionInAlignment + i);
797 bases << (ba.IsReverseStrand() ? (char)tolower(insertedBase) : (char)toupper(insertedBase) );
801 // if next position contains DELETION
802 else if ( pa.IsNextDeletion ) {
803 bases << '-' << pa.DeletionLength;
804 for (int i = 1; i <= pa.DeletionLength; ++i) {
805 char deletedBase('N');
806 if ( m_hasFasta && (pileupData.Position+i < m_references[pileupData.RefId].RefLength) ) {
807 if ( !m_fasta.GetBase(pileupData.RefId, pileupData.Position+i, deletedBase ) ) {
808 cerr << "Pileup error : Could not read reference base from FASTA file" << endl;
812 bases << (ba.IsReverseStrand() ? (char)tolower(deletedBase) : (char)toupper(deletedBase) );
817 // otherwise, DELETION
820 // if end of read segment
821 if ( pa.IsSegmentEnd ) bases << '$';
823 // store current base quality
824 baseQualities << ba.Qualities.at(pa.PositionInAlignment);
826 // save alignment map quality if desired
827 if ( m_isPrintingMapQualities )
828 mapQualities << ( ((int)ba.MapQuality > 93) ? (char)126 : (char)((int)ba.MapQuality+33) );
831 // ----------------------
835 // <refName> <1-based pos> <refBase> <numberAlleles> <bases> <qualities> [mapQuals]
837 const string TAB = "\t";
838 *m_out << referenceName << TAB
839 << position + 1 << TAB
840 << referenceBase << TAB
841 << numberAlleles << TAB
842 << bases.str() << TAB
843 << baseQualities.str() << TAB
844 << mapQualities.str() << endl;