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/*
* =====================================================================================
*
* Filename: SAMFilter.cpp
*
* Description: Filter SAM Hits according to
* filteration criteria
* minPctSimilarity, minAccuracy,
* minLength, holeNumbers
* and multiple-hit policy
* random : a random hit
* all : all hits
* allbest : all hits with the best score
* randombest: a random hit selected from all the hits
* that have the best score
*
* Version: 1.0
* Created: 03/19/2013 01:19:43 PM
* Revision: none
* Compiler: gcc
*
* Author: Yuan Li (yli), yli@pacificbiosciences.com
* Company: Pacific Biosciences
*
* =====================================================================================
*/
#include <climits>
#include <iostream>
#include <pbdata/defs.h>
#include <alignment/algorithms/alignment/AlignmentUtils.hpp>
#include <alignment/algorithms/alignment/DistanceMatrixScoreFunction.hpp>
#include <alignment/algorithms/alignment/StringToScoreMatrix.hpp>
#include <alignment/datastructures/alignment/AlignmentCandidate.hpp>
#include <alignment/datastructures/alignment/FilterCriteria.hpp>
#include <alignment/datastructures/alignment/SAMToAlignmentCandidateAdapter.hpp>
#include <alignment/format/SAMPrinter.hpp>
#include <alignment/utils/RangeUtils.hpp>
#include <pbdata/ChangeListID.hpp>
#include <pbdata/CommandLineParser.hpp>
#include <pbdata/FASTAReader.hpp>
#include <pbdata/FASTASequence.hpp>
#include <pbdata/GFFFile.hpp>
#include <pbdata/metagenome/TitleTable.hpp>
#include <pbdata/sam/SAMReader.hpp>
#include <pbdata/utils/SMRTReadUtils.hpp>
#include <pbdata/utils/TimeUtils.hpp>
#include "../iblasr/RegisterFilterOptions.h"
//#define USE_GOOGLE_PROFILER
#ifdef USE_GOOGLE_PROFILER
#include <gperftools/profiler.h>
#endif
char VERSION[] = "v0.1.0";
char PERFORCE_VERSION_STRING[] = "$Change: 134995 $";
// By default negative score is better.
ScoreSign scoreSign = ScoreSign::NEGATIVE;
// Compare SAMAlignment objects by qName, score and
// target positions.
bool byQNameScoreTStart(const SAMAlignment &a, const SAMAlignment &b)
{
if (a.qName == b.qName) {
if (a.score == b.score) return a.pos < b.pos;
return Score(a.score, scoreSign).WorseThan(Score(b.score, scoreSign));
}
return (a.qName < b.qName);
}
// Compare SAMAlignment objects by rName and qName
bool byRNameQName(const SAMAlignment &a, const SAMAlignment &b)
{
if (a.rName == b.rName) {
return a.qName < b.qName;
}
return (a.rName < b.rName);
}
// Get the next group of SAM alignments that have the same qName from
// allSAMAlignments[groupBegin ... groupEnd)
// Note that allSAMAlignments is already sorted by qName, score and tPos.
void GetNextSAMAlignmentGroup(std::vector<SAMAlignment> &allSAMAlignments, unsigned int groupBegin,
unsigned int &groupEnd)
{
assert(groupBegin < allSAMAlignments.size());
groupEnd = groupBegin + 1;
std::string queryName = allSAMAlignments[groupBegin].qName;
while (groupEnd < allSAMAlignments.size()) {
if (allSAMAlignments[groupEnd].qName == queryName)
groupEnd++;
else
break;
}
}
// Get the best SAM alignments whose alignment score are the best.
// Assume that alignments in allSAMAlignments[groupBegin, groupEnd)
// all have the same queryName and are sorted by score and tPos
// asscendingly: worst, ...., best
void GetBestSAMAlignmentsInGroup(std::vector<SAMAlignment> &allSAMAlignments,
const unsigned int &groupBegin, const unsigned int &groupEnd,
unsigned int &bestBegin, unsigned int &bestEnd)
{
assert(groupEnd <= allSAMAlignments.size() and groupBegin < groupEnd);
bestEnd = groupEnd;
bestBegin = groupEnd - 1;
int groupBestScore = allSAMAlignments[bestBegin].score;
std::string queryName = allSAMAlignments[bestBegin].qName;
while (bestBegin >= groupBegin and bestBegin < groupEnd) {
assert(allSAMAlignments[bestBegin].qName == queryName);
if (allSAMAlignments[bestBegin].score == groupBestScore)
bestBegin -= 1;
else
break;
}
bestBegin += 1;
}
// Apply hit policy to a group of SAM alignments and return indices
// of the selected alignments.
std::vector<unsigned int> ApplyHitPolicy(HitPolicy &hitPolicy,
std::vector<SAMAlignment> &allSAMAlignments,
const unsigned int &groupBegin,
const unsigned int &groupEnd)
{
std::vector<unsigned int> hitIndices;
if (hitPolicy.IsAll()) {
for (unsigned int i = groupBegin; i < groupEnd; i++) {
hitIndices.push_back(i);
}
} else if (hitPolicy.IsRandom()) {
hitIndices.push_back(rand() % (groupEnd - groupBegin) + groupBegin);
} else {
unsigned int bestBegin, bestEnd;
GetBestSAMAlignmentsInGroup(allSAMAlignments, groupBegin, groupEnd, bestBegin, bestEnd);
if (hitPolicy.IsAllbest()) {
for (unsigned int i = bestBegin; i < bestEnd; i++) {
hitIndices.push_back(i);
}
} else if (hitPolicy.IsRandombest()) {
hitIndices.push_back(rand() % (bestEnd - bestBegin) + bestBegin);
} else if (hitPolicy.IsLeftmost()) {
hitIndices.push_back(bestBegin);
} else {
assert(false);
}
}
return hitIndices;
}
// Convert references[...].title in reference.fasta to their corresponding
// indices in the title table.
void ConvertTitlesToTitleTableIndices(std::vector<FASTASequence> &references,
std::string &titleTableName)
{
TitleTable tt;
tt.Read(titleTableName);
for (size_t i = 0; i < references.size(); i++) {
std::string title = references[i].GetTitle();
int idx = -1;
if (tt.Lookup(title, idx)) {
std::stringstream ss;
ss << idx;
references[i].CopyTitle(ss.str());
} else {
std::cout << "ERROR, reference " << title << " does not exist "
<< " in the title table " << titleTableName << ". The "
<< "reference fasta and the title table do not match." << std::endl;
std::exit(EXIT_FAILURE);
}
}
tt.Free();
}
// Return true if the alignment can only map to an adapter specified
// in the adapter GFF file.
// A sample record in adapter GFF file:
// ref000001 . adapter 10955 10999 0.00 + . xxxx
// ref000001 . adapter 32886 32930 0.00 + . xxxx
// Note that the first field (e.g., 'ref000001') is id of sequence
// in a reference repository, not sequence name, so we need to
// reconstruct the mapping between sequence id and sequence name.
bool CheckAdapterOnly(GFFFile &adapterGffFile, //Adapter gff file
AlignmentCandidate<> &alignment, // An alignment
std::map<std::string, int> &refNameToIndex)
{
// Map target sequence name to its index in reference repository.
if (refNameToIndex.find(alignment.tName) == refNameToIndex.end()) {
// This should not happen ...
std::cout << "ERROR, could not find alignment target name " << alignment.tName
<< " in the reference file." << std::endl;
std::exit(EXIT_FAILURE);
}
int refNameIndex = refNameToIndex[alignment.tName];
char buf[16];
sprintf(buf, "ref%06d", refNameIndex + 1);
// Reconstruct ref id in the format "ref00000?".
std::string refNameId(buf);
int FUZZY_OVERLAP = 20;
for (size_t eindex = 0; eindex < adapterGffFile.entries.size(); eindex++) {
GFFEntry &entry = adapterGffFile.entries[eindex];
// Convert each GFF record from 1-based inclusive to
// 0-based exclusive.
if (entry.type == "adapter" and
(entry.name == alignment.tName or entry.name == refNameId)) {
UInt estart = entry.start - 1;
UInt eend = entry.end;
if (entry.strand == '-') {
UInt tmp = estart;
estart = alignment.tLength - 1 - eend;
eend = alignment.tLength - 1 - tmp;
}
if (not(eend < alignment.GenomicTBegin() or estart > alignment.GenomicTEnd())) {
UInt lengthUnion = std::max(eend, alignment.GenomicTEnd()) -
std::min(estart, alignment.GenomicTBegin());
if (lengthUnion < eend - estart + FUZZY_OVERLAP) {
return true;
}
}
}
}
return false;
}
int main(int argc, char *argv[])
{
#ifdef USE_GOOGLE_PROFILER
char *profileFileName = getenv("CPUPROFILE");
if (profileFileName != NULL) {
ProfilerStart(profileFileName);
} else {
ProfilerStart("google_profile.txt");
}
#endif
// Register inputs and outputs.
std::string samFileName, refFileName, outFileName;
CommandLineParser clp;
clp.RegisterStringOption("file.sam", &samFileName, "Input SAM file.");
clp.RegisterStringOption("reference.fasta", &refFileName, "Reference used to generate reads.");
clp.RegisterStringOption("out.sam", &outFileName, "Output SAM file.");
clp.RegisterPreviousFlagsAsHidden();
// Register filter criteria options.
int minAlnLength = 50;
float minPctSimilarity = 70, minPctAccuracy = 70;
std::string hitPolicyStr = "randombest";
bool useScoreCutoff = false;
int scoreCutoff = INF_INT;
int scoreSignInt = -1;
RegisterFilterOptions(clp, minAlnLength, minPctSimilarity, minPctAccuracy, hitPolicyStr,
useScoreCutoff, scoreSignInt, scoreCutoff);
int seed = 1;
clp.RegisterIntOption("seed", &seed,
"(1) Seed for random number generator.\n"
"If seed is 0, then use current time as seed.",
CommandLineParser::Integer);
std::string holeNumberStr;
Ranges holeNumberRanges;
clp.RegisterStringOption("-holeNumbers", &holeNumberStr,
"A string of comma-delimited hole number ranges to output hits, "
"such as '1,2,10-12'. "
"This requires hit titles to be in SMRT read title format.");
bool parseSmrtTitle = false;
clp.RegisterFlagOption("smrtTitle", &parseSmrtTitle,
"Use this option when filtering alignments generated by "
"programs other than blasr, e.g. bwa-sw or gmap. "
" Parse read coordinates from the SMRT read title. "
"The title is in the format /name/hole/coordinates, where"
" coordinates are in the format \\d+_\\d+, and represent "
"the interval of the read that was aligned.");
/* This experimental option can be useful for metagenomics, in which case
* there are hundreds of sequences in the target, of which many titles are
* long and may contain white spaces (e.g., ' ', '\t').
* In order to save disc space and avoid the (possibly) none unique mapping
* between full and short reference names, one may call blasr with
* -titleTable option to represent all target sequences in the output
* by their indices in the title table.*/
std::string titleTableName = "";
clp.RegisterStringOption("titleTable", &titleTableName,
"Use this experimental option when filtering alignments generated by "
"blasr with -titleTable titleTableName, in which case "
"reference titles in SAM are represented by their "
"indices (e.g., 0, 1, 2, ...) in the title table.");
std::string adapterGffFileName = "";
clp.RegisterStringOption("filterAdapterOnly", &adapterGffFileName,
"Use this option to remove reads which can only map to adapters "
"specified in the GFF file.");
bool verbose = false;
clp.RegisterFlagOption("v", &verbose, "Be verbose.");
clp.SetExamples(
"Because SAM has optional tags that have different meanings"
" in different programs, careful usage is required in order "
"to have proper output. The \"xs\" tag in bwa-sw is used to "
"show the suboptimal score, but in PacBio SAM (blasr) it is "
"defined as the start in the query sequence of the alignment.\n"
"When \"-smrtTitle\" is specified, the xs tag is ignored, but "
"when it is not specified, the coordinates given by the xs and "
"xe tags are used to define the interval of a read that is "
"aligned. The CIGAR string is relative to this interval.");
clp.ParseCommandLine(argc, argv);
// Set random number seed.
if (seed == 0) {
srand(time(NULL));
} else {
srand(seed);
}
scoreSign = (scoreSignInt == -1) ? ScoreSign::NEGATIVE : ScoreSign::POSITIVE;
Score s(static_cast<float>(scoreCutoff), scoreSign);
FilterCriteria filterCriteria(minAlnLength, minPctSimilarity, minPctAccuracy, true, s);
filterCriteria.Verbose(verbose);
HitPolicy hitPolicy(hitPolicyStr, scoreSign);
std::string errMsg;
if (not filterCriteria.MakeSane(errMsg)) {
std::cout << errMsg << std::endl;
std::exit(EXIT_FAILURE);
}
// Parse hole number ranges.
if (holeNumberStr.size() != 0) {
if (not holeNumberRanges.setRanges(holeNumberStr)) {
std::cout << "Could not parse hole number ranges: " << holeNumberStr << "."
<< std::endl;
std::exit(EXIT_FAILURE);
}
}
// Open output file.
std::ofstream outFileStrm;
if (outFileName != "") {
CrucialOpen(outFileName, outFileStrm, std::ios::out);
}
GFFFile adapterGffFile;
if (adapterGffFileName != "") adapterGffFile.ReadAll(adapterGffFileName);
SAMReader<SAMFullReferenceSequence, SAMReadGroup, SAMAlignment> samReader;
FASTAReader fastaReader;
//
// Initialize samReader and fastaReader.
//
samReader.Initialize(samFileName);
fastaReader.Initialize(refFileName);
//
// Configure the file log.
//
std::string command;
CommandLineParser::CommandLineToString(argc, argv, command);
std::string log = "Filter sam hits.";
std::string program = "samFilter";
std::string versionString = VERSION;
AppendPerforceChangelist(PERFORCE_VERSION_STRING, versionString);
//
// Read necessary input.
//
std::vector<FASTASequence> references;
fastaReader.ReadAllSequences(references);
// If the SAM file is generated by blasr with -titleTable,
// then references in the SAM are represented by
// their corresponding indices in the title table.
// In that case, we need to convert reference titles in fasta file
// to their corresponding indices in the title table, such that
// references in both SAM and fasta files are represented
// by title table indices and therefore can match.
if (titleTableName != "") {
ConvertTitlesToTitleTableIndices(references, titleTableName);
}
AlignmentSet<SAMFullReferenceSequence, SAMReadGroup, SAMAlignment> alignmentSet;
std::vector<std::string> allHeaders = samReader.ReadHeader(alignmentSet);
// Process SAM Header.
std::string commandLineString;
clp.CommandLineToString(argc, argv, commandLineString);
allHeaders.push_back("@PG\tID:SAMFILTER\tVN:" + versionString + "\tCL:" + program + " " +
commandLineString);
for (size_t i = 0; i < allHeaders.size(); i++) {
outFileStrm << allHeaders[i] << std::endl;
}
//
// The order of references in std::vector<FASTASequence> references and
// AlignmentSet<, , >alignmentSet.references can be different.
// Rearrange alignmentSet.references such that they are ordered in
// exactly the same way as std::vector<FASTASequence> references.
//
alignmentSet.RearrangeReferences(references);
// Map reference name obtained from SAM file to indices
std::map<std::string, int> refNameToIndex;
for (size_t i = 0; i < references.size(); i++) {
std::string refName = alignmentSet.references[i].GetSequenceName();
refNameToIndex[refName] = i;
}
//
// Store the alignments.
//
SAMAlignment samAlignment;
size_t alignIndex = 0;
//
// For 150K, each chip produces about 300M sequences
// (not including quality values and etc.).
// Let's assume that the sam file and reference data can
// fit in the memory.
// Need to scale for larger sequal data in the future.
//
std::vector<SAMAlignment> allSAMAlignments;
while (samReader.GetNextAlignment(samAlignment)) {
if (samAlignment.rName == "*") {
continue;
}
if (parseSmrtTitle and holeNumberStr.size() != 0) {
std::string movieName;
int thisHoleNumber;
if (not ParsePBIReadName(samAlignment.qName, movieName, thisHoleNumber)) {
std::cout << "ERROR, could not parse SMRT title: " << samAlignment.qName << "."
<< std::endl;
std::exit(EXIT_FAILURE);
}
if (not holeNumberRanges.contains(UInt(thisHoleNumber))) {
if (verbose) std::cout << thisHoleNumber << " is not in range." << std::endl;
continue;
}
}
if (samAlignment.cigar.find('P') != std::string::npos) {
std::cout << "WARNING. Could not process SAM record with 'P' in "
<< "its cigar string." << std::endl;
continue;
}
std::vector<AlignmentCandidate<> > convertedAlignments;
SAMAlignmentsToCandidates(samAlignment, references, refNameToIndex, convertedAlignments,
parseSmrtTitle, false);
if (convertedAlignments.size() > 1) {
std::cout << "WARNING. Ignore multiple segments." << std::endl;
continue;
}
for (int i = 0; i < 1; i++) {
AlignmentCandidate<> &alignment = convertedAlignments[i];
//score func does not matter
DistanceMatrixScoreFunction<DNASequence, DNASequence> distFunc;
ComputeAlignmentStats(alignment, alignment.qAlignedSeq.seq, alignment.tAlignedSeq.seq,
distFunc);
// Check whether this alignment can only map to adapters in
// the adapter GFF file.
if (adapterGffFileName != "" and
CheckAdapterOnly(adapterGffFile, alignment, refNameToIndex)) {
if (verbose) std::cout << alignment.qName << " filter adapter only." << std::endl;
continue;
}
// Assign score to samAlignment.
samAlignment.score = samAlignment.as;
if (not filterCriteria.Satisfy(static_cast<AlignmentCandidate<> *>(&alignment))) {
continue;
}
allSAMAlignments.push_back(samAlignment);
alignment.FreeSubsequences();
}
++alignIndex;
}
// Sort all SAM alignments by qName, score and target position.
sort(allSAMAlignments.begin(), allSAMAlignments.end(), byQNameScoreTStart);
unsigned int groupBegin = 0;
unsigned int groupEnd = -1;
std::vector<SAMAlignment> filteredSAMAlignments;
while (groupBegin < allSAMAlignments.size()) {
// Get the next group of SAM alignments which have the same qName
// from allSAMAlignments[groupBegin ... groupEnd)
GetNextSAMAlignmentGroup(allSAMAlignments, groupBegin, groupEnd);
std::vector<unsigned int> hitIndices =
ApplyHitPolicy(hitPolicy, allSAMAlignments, groupBegin, groupEnd);
for (unsigned int i = 0; i < hitIndices.size(); i++) {
filteredSAMAlignments.push_back(allSAMAlignments[hitIndices[i]]);
}
groupBegin = groupEnd;
}
// Sort all SAM alignments by reference name and query name
sort(filteredSAMAlignments.begin(), filteredSAMAlignments.end(), byRNameQName);
for (unsigned int i = 0; i < filteredSAMAlignments.size(); i++) {
filteredSAMAlignments[i].PrintSAMAlignment(outFileStrm);
}
if (outFileName != "") {
outFileStrm.close();
}
#ifdef USE_GOOGLE_PROFILER
ProfilerStop();
#endif
return 0;
}
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