コード例 #1
0
ファイル: Stats.cpp プロジェクト: BioScripts/bamUtil
int Stats::execute(int argc, char **argv)
{
    // Extract command line arguments.
    String inFile = "";
    String indexFile = "";
    bool basic = false;
    bool noeof = false;
    bool params = false;
    bool qual = false;
    bool phred = false;
    int maxNumReads = -1;
    bool unmapped = false;
    String pBaseQC = "";
    String cBaseQC = "";
    String regionList = "";
    int excludeFlags = 0;
    int requiredFlags = 0;
    bool withinRegion = false;
    int minMapQual = 0;
    String dbsnp = "";
    PosList *dbsnpListPtr = NULL;
    bool baseSum = false;
    int bufferSize = PileupHelper::DEFAULT_WINDOW_SIZE;

    ParameterList inputParameters;
    BEGIN_LONG_PARAMETERS(longParameterList)
        LONG_PARAMETER_GROUP("Required Parameters")
        LONG_STRINGPARAMETER("in", &inFile)
        LONG_PARAMETER_GROUP("Types of Statistics")
        LONG_PARAMETER("basic", &basic)
        LONG_PARAMETER("qual", &qual)
        LONG_PARAMETER("phred", &phred)
        LONG_STRINGPARAMETER("pBaseQC", &pBaseQC)
        LONG_STRINGPARAMETER("cBaseQC", &cBaseQC)
        LONG_PARAMETER_GROUP("Optional Parameters")
        LONG_INTPARAMETER("maxNumReads", &maxNumReads)
        LONG_PARAMETER("unmapped", &unmapped)
        LONG_STRINGPARAMETER("bamIndex", &indexFile)
        LONG_STRINGPARAMETER("regionList", &regionList)
        LONG_INTPARAMETER("excludeFlags", &excludeFlags)
        LONG_INTPARAMETER("requiredFlags", &requiredFlags)
        LONG_PARAMETER("noeof", &noeof)
        LONG_PARAMETER("params", &params)
        LONG_PARAMETER_GROUP("Optional phred/qual Only Parameters")
        LONG_PARAMETER("withinRegion", &withinRegion)
        LONG_PARAMETER_GROUP("Optional BaseQC Only Parameters")
        LONG_PARAMETER("baseSum", &baseSum)
        LONG_INTPARAMETER("bufferSize", &bufferSize)
        LONG_INTPARAMETER("minMapQual", &minMapQual)
        LONG_STRINGPARAMETER("dbsnp", &dbsnp)
        END_LONG_PARAMETERS();
   
    inputParameters.Add(new LongParameters ("Input Parameters", 
                                            longParameterList));

    inputParameters.Read(argc-1, &(argv[1]));

    // If no eof block is required for a bgzf file, set the bgzf file type to 
    // not look for it.
    if(noeof)
    {
        // Set that the eof block is not required.
        BgzfFileType::setRequireEofBlock(false);
    }

    // Check to see if the in file was specified, if not, report an error.
    if(inFile == "")
    {
        usage();
        inputParameters.Status();
        // In file was not specified but it is mandatory.
        std::cerr << "--in is a mandatory argument for stats, "
                  << "but was not specified" << std::endl;
        return(-1);
    }

    // Use the index file if unmapped or regionList is not empty.
    bool useIndex = (unmapped|| (!regionList.IsEmpty()));

    // IndexFile is required, so check to see if it has been set.
    if(useIndex && (indexFile == ""))
    {
        // In file was not specified, so set it to the in file
        // + ".bai"
        indexFile = inFile + ".bai";
    }
    ////////////////////////////////////////
    // Setup in case pileup is used.
    Pileup<PileupElementBaseQCStats> pileup(bufferSize);
    // Initialize start/end positions.
    myStartPos = 0;
    myEndPos = -1;
    
    // Open the output qc file if applicable.
    IFILE baseQCPtr = NULL;
    if(!pBaseQC.IsEmpty() && !cBaseQC.IsEmpty())
    {
        usage();
        inputParameters.Status();
        // Cannot specify both types of baseQC.
        std::cerr << "Cannot specify both --pBaseQC & --cBaseQC." << std::endl;
        return(-1);
    }
    else if(!pBaseQC.IsEmpty())
    {
        baseQCPtr = ifopen(pBaseQC, "w");
        PileupElementBaseQCStats::setPercentStats(true);
    }
    else if(!cBaseQC.IsEmpty())
    {
        baseQCPtr = ifopen(cBaseQC, "w");
        PileupElementBaseQCStats::setPercentStats(false);
    }

    if(baseQCPtr != NULL)
    {
        PileupElementBaseQCStats::setOutputFile(baseQCPtr);
        PileupElementBaseQCStats::printHeader();
    }
    if((baseQCPtr != NULL) || baseSum)
    {
        PileupElementBaseQCStats::setMapQualFilter(minMapQual);
        PileupElementBaseQCStats::setBaseSum(baseSum);
    }

    if(params)
    {
        inputParameters.Status();
    }

    // Open the file for reading.
    SamFile samIn;
    if(!samIn.OpenForRead(inFile))
    {
        fprintf(stderr, "%s\n", samIn.GetStatusMessage());
        return(samIn.GetStatus());
    }

    samIn.SetReadFlags(requiredFlags, excludeFlags);

    // Set whether or not basic statistics should be generated.
    samIn.GenerateStatistics(basic);

    // Read the sam header.
    SamFileHeader samHeader;
    if(!samIn.ReadHeader(samHeader))
    {
        fprintf(stderr, "%s\n", samIn.GetStatusMessage());
        return(samIn.GetStatus());
    }

    // Open the bam index file for reading if we are
    // doing unmapped reads (also set the read section).
    if(useIndex)
    {
        samIn.ReadBamIndex(indexFile);

        if(unmapped)
        {
            samIn.SetReadSection(-1);
        }

        if(!regionList.IsEmpty())
        {
            myRegionList = ifopen(regionList, "r");
        }
    }

    //////////////////////////
    // Read dbsnp if specified and doing baseQC
    if(((baseQCPtr != NULL) || baseSum) && (!dbsnp.IsEmpty()))
    {
        // Read the dbsnp file.
        IFILE fdbSnp;
        fdbSnp = ifopen(dbsnp,"r");
        // Determine how many entries.
        const SamReferenceInfo& refInfo = samHeader.getReferenceInfo();
        int maxRefLen = 0;
        for(int i = 0; i < refInfo.getNumEntries(); i++)
        {
            int refLen = refInfo.getReferenceLength(i);
            if(refLen >= maxRefLen)
            {
                maxRefLen = refLen + 1;
            }
        }
        
        dbsnpListPtr = new PosList(refInfo.getNumEntries(),maxRefLen);

        if(fdbSnp==NULL)
        {
            std::cerr << "Open dbSNP file " << dbsnp.c_str() << " failed!\n";
        }
        else if(dbsnpListPtr == NULL)
        {
            std::cerr << "Failed to init the memory allocation for the dbsnpList.\n";
        }
        else
        {
            // Read the dbsnp file.
            StringArray tokens;
            String buffer;
            int position = 0;
            int refID = 0;

            // Loop til the end of the file.
            while (!ifeof(fdbSnp))
            {
                // Read the next line.
                buffer.ReadLine(fdbSnp);
                // If it does not have at least 2 columns, 
                // continue to the next line.
                if (buffer.IsEmpty() || buffer[0] == '#') continue;
                tokens.AddTokens(buffer);
                if(tokens.Length() < 2) continue;

                if(!tokens[1].AsInteger(position))
                {
                    std::cerr << "Improperly formatted region line, start position "
                              << "(2nd column) is not an integer: "
                              << tokens[1]
                              << "; Skipping to the next line.\n";         
                    continue;
                }

                // Look up the reference name.
                refID = samHeader.getReferenceID(tokens[0]);
                if(refID != SamReferenceInfo::NO_REF_ID)
                {
                    // Reference id was found, so add it to the dbsnp
                    dbsnpListPtr->addPosition(refID, position);
                }
        
                tokens.Clear();
                buffer.Clear();
            }
        }
        ifclose(fdbSnp);
    }

    // Read the sam records.
    SamRecord samRecord;

    int numReads = 0;

    //////////////////////
    // Setup in case doing a quality count.
    // Quality histogram.
    const int MAX_QUAL = 126;
    const int START_QUAL = 33;
    uint64_t qualCount[MAX_QUAL+1];
    for(int i = 0; i <= MAX_QUAL; i++)
    {
        qualCount[i] = 0;
    }
    
    const int START_PHRED = 0;
    const int PHRED_DIFF = START_QUAL - START_PHRED;
    const int MAX_PHRED = MAX_QUAL - PHRED_DIFF;
    uint64_t phredCount[MAX_PHRED+1];
    for(int i = 0; i <= MAX_PHRED; i++)
    {
        phredCount[i] = 0;
    }
    
    int refPos = 0;
    Cigar* cigarPtr = NULL;
    char cigarChar = '?';
    // Exclude clips from the qual/phred counts if unmapped reads are excluded.
    bool qualExcludeClips = excludeFlags & SamFlag::UNMAPPED;

    //////////////////////////////////
    // When not reading by sections, getNextSection returns true
    // the first time, then false the next time.
    while(getNextSection(samIn))
    {
        // Keep reading records from the file until SamFile::ReadRecord
        // indicates to stop (returns false).
        while(((maxNumReads < 0) || (numReads < maxNumReads)) && samIn.ReadRecord(samHeader, samRecord))
        {
            // Another record was read, so increment the number of reads.
            ++numReads;
            // See if the quality histogram should be genereated.
            if(qual || phred)
            {
                // Get the quality.
                const char* qual = samRecord.getQuality();
                // Check for no quality ('*').
                if((qual[0] == '*') && (qual[1] == 0))
                {
                    // This record does not have a quality string, so no 
                    // quality processing is necessary.
                }
                else
                {
                    int index = 0;
                    cigarPtr = samRecord.getCigarInfo();
                    cigarChar = '?';
                    refPos = samRecord.get0BasedPosition();
                    if(!qualExcludeClips && (cigarPtr != NULL))
                    {
                        // Offset the reference position by any soft clips
                        // by subtracting the queryIndex of this start position.
                        // refPos is now the start position of the clips.
                        refPos -= cigarPtr->getQueryIndex(0);
                    }

                    while(qual[index] != 0)
                    {
                        // Skip this quality if it is clipped and we are skipping clips.
                        if(cigarPtr != NULL)
                        {
                            cigarChar = cigarPtr->getCigarCharOpFromQueryIndex(index);
                        }
                        if(qualExcludeClips && Cigar::isClip(cigarChar))
                        {
                            // Skip a clipped quality.
                            ++index;
                            // Increment the position.
                            continue;
                        }

                        if(withinRegion && (myEndPos != -1) && (refPos >= myEndPos))
                        {
                            // We have hit the end of the region, stop processing this
                            // quality string.
                            break;
                        }

                        if(withinRegion && (refPos < myStartPos))
                        {
                            // This position is not in the target.
                            ++index;
                            // Update the position if this is found in the reference or a clip.
                            if(Cigar::foundInReference(cigarChar) || Cigar::isClip(cigarChar))
                            {
                                ++refPos;
                            }
                            continue;
                        }

                        // Check for valid quality.
                        if((qual[index] < START_QUAL) || (qual[index] > MAX_QUAL))
                        {
                            if(qual)
                            {
                                std::cerr << "Invalid Quality found: " << qual[index] 
                                          << ".  Must be between "
                                          << START_QUAL << " and " << MAX_QUAL << ".\n";
                            }
                            if(phred)
                            {
                                std::cerr << "Invalid Phred Quality found: " << qual[index] - PHRED_DIFF
                                          << ".  Must be between "
                                          << START_QUAL << " and " << MAX_QUAL << ".\n";
                            }
                            // Skip an invalid quality.
                            ++index;
                            // Update the position if this is found in the reference or a clip.
                            if(Cigar::foundInReference(cigarChar) || Cigar::isClip(cigarChar))
                            {
                                ++refPos;
                            }
                            continue;
                        }
                        
                        // Increment the count for this quality.
                        ++(qualCount[(int)(qual[index])]);
                        ++(phredCount[(int)(qual[index]) - PHRED_DIFF]);
                        // Update the position if this is found in the reference or a clip.
                        if(Cigar::foundInReference(cigarChar) || Cigar::isClip(cigarChar))
                        {
                            ++refPos;
                        }
                        ++index;
                    }
                }
            }

            // Check the next thing to do for the read.
            if((baseQCPtr != NULL) || baseSum)
            {
                // Pileup the bases for this read.
                pileup.processAlignmentRegion(samRecord, myStartPos, myEndPos, dbsnpListPtr);
            }
        }

        // Done with a section, move on to the next one.

        // New section, so flush the pileup.
        pileup.flushPileup();
    }

    // Flush the rest of the pileup.
    if((baseQCPtr != NULL) || baseSum)
    {
        // Pileup the bases.
        pileup.processAlignmentRegion(samRecord, myStartPos, myEndPos, dbsnpListPtr);
        PileupElementBaseQCStats::printSummary();
        ifclose(baseQCPtr);
    }

    std::cerr << "Number of records read = " << 
        samIn.GetCurrentRecordCount() << std::endl;

    if(basic)
    {
        std::cerr << std::endl;
        samIn.PrintStatistics();
    }

    // Print the quality stats.
    if(qual)
    {
        std::cerr << std::endl;
        std::cerr << "Quality\tCount\n";
        for(int i = START_QUAL; i <= MAX_QUAL; i++)
        {
            std::cerr << i << "\t" << qualCount[i] << std::endl;
        }
    }
    // Print the phred quality stats.
    if(phred)
    {
        std::cerr << std::endl;
        std::cerr << "Phred\tCount\n";
        for(int i = START_PHRED; i <= MAX_PHRED; i++)
        {
            std::cerr << i << "\t" << phredCount[i] << std::endl;
        }
    }

    SamStatus::Status status = samIn.GetStatus();
    if(status == SamStatus::NO_MORE_RECS)
    {
        // A status of NO_MORE_RECS means that all reads were successful.
        status = SamStatus::SUCCESS;
    }

    return(status);
}
コード例 #2
0
// Add an entry to this pileup element.  
void PileupElementBaseQual::addEntry(SamRecord& record)
{
    // Call the base class:
    PileupElement::addEntry(record);

    if(myRefAllele.empty())
    {
    	genomeIndex_t markerIndex = (*myRefSeq).getGenomePosition(getChromosome(), static_cast<uint32_t>(getRefPosition()+1));
        myRefAllele = (*myRefSeq)[markerIndex];
    }

    // Increment the index
    ++myIndex;
    
    // if the index has gone beyond the allocated space, double the size.
    if(myIndex >= myAllocatedSize)
    {
        char* tempBuffer = (char*)realloc(myBases, myAllocatedSize * 2);
        if(tempBuffer == NULL)
        {
            std::cerr << "Memory Allocation Failure\n";
            // TODO
            return;
        }
        myBases = tempBuffer;
        int8_t* tempInt8Buffer = (int8_t*)realloc(myMapQualities, myAllocatedSize * 2 * sizeof(int8_t));
        if(tempInt8Buffer == NULL)
        {
            std::cerr << "Memory Allocation Failure\n";
            // TODO
            return;
        }
        myMapQualities = tempInt8Buffer; 
        tempInt8Buffer = (int8_t*)realloc(myQualities, myAllocatedSize * 2 * sizeof(int8_t));
        if(tempInt8Buffer == NULL)
        {
            std::cerr << "Memory Allocation Failure\n";
            // TODO
            return;
        }
        myQualities = tempInt8Buffer;
        tempBuffer = (char*)realloc(myStrands, myAllocatedSize * 2);
        if(tempBuffer == NULL)
        {
            std::cerr << "Memory Allocation Failure\n";
            // TODO
            return;
        }
        myStrands = tempBuffer;
        tempInt8Buffer = (int8_t*)realloc(myCycles, myAllocatedSize * 2 * sizeof(int8_t));
        if(tempInt8Buffer == NULL)
        {
            std::cerr << "Memory Allocation Failure\n";
            // TODO
            return;
        }
        myCycles = tempInt8Buffer; 
        int16_t* tempInt16Buffer = (int16_t*)realloc(myGLScores, myAllocatedSize * 2 * sizeof(int16_t));
        if(tempInt8Buffer == NULL)
        {
            std::cerr << "Memory Allocation Failure\n";
            // TODO
            return;
        }
        myGLScores = tempInt16Buffer;
        myAllocatedSize = myAllocatedSize * 2;
    }

    Cigar* cigar = record.getCigarInfo();
    
    if(cigar == NULL)
    {
        throw std::runtime_error("Failed to retrieve cigar info from the record.");
    }

    int32_t readIndex = 
        cigar->getQueryIndex(getRefPosition(), record.get0BasedPosition());

    // If the readPosition is N/A, this is a deletion.
    if(readIndex != CigarRoller::INDEX_NA)
    {
        char base = record.getSequence(readIndex);
        int8_t mapQual = record.getMapQuality();
        //-33 to obtain the PHRED base quality
        char qual = record.getQuality(readIndex) - 33;
        if(qual == UNSET_QUAL)
        {
            qual = ' ';
        }
        char strand = (record.getFlag() & 0x0010) ? 'R' : 'F';
        int cycle = strand == 'F' ? readIndex + 1 : record.getReadLength() -  readIndex;
        myBases[myIndex] = base;
        myMapQualities[myIndex] = mapQual;
        myQualities[myIndex] = qual;
        myStrands[myIndex] = strand;
        myCycles[myIndex] = cycle;
    }
    else if(myAddDelAsBase)
    {
        int8_t mapQual = record.getMapQuality();
        char strand = (record.getFlag() & 0x0010) ? 'R' : 'F';
        myBases[myIndex] = '-';
        myMapQualities[myIndex] = mapQual;
        myQualities[myIndex] = -1;
        myStrands[myIndex] = strand;
        myCycles[myIndex] = -1;
    }
    else
    {
        // Do not add a deletion.
        // Did not add any entries, so decrement the index counter since the
        // index was not used.
        --myIndex;
    }
}