package org.broadinstitute.sting.alignment; import org.broadinstitute.sting.gatk.walkers.ReadWalker; import org.broadinstitute.sting.utils.BaseUtils; import org.broadinstitute.sting.utils.StingException; import org.broadinstitute.sting.utils.cmdLine.Argument; import org.broadinstitute.sting.alignment.Alignment; import org.broadinstitute.sting.alignment.bwa.c.BWACAligner; import org.broadinstitute.sting.alignment.bwa.c.BWACConfiguration; import net.sf.samtools.SAMRecord; /** * Validates alignments against existing reads. * * @author mhanna * @version 0.1 */ public class AlignmentValidationWalker extends ReadWalker { /** * The supporting BWT index generated using BWT. */ @Argument(fullName="BWTPrefix",shortName="BWT",doc="Index files generated by bwa index -d bwtsw",required=false) String prefix = "/Users/mhanna/reference/Ecoli/Escherichia_coli_K12_MG1655.fasta"; /** * The instance used to generate alignments. */ private BWACAligner aligner = null; /** * Number of reads processed. */ private int count = 0; /** * Max delta in mapping quality. */ private int maxMapQDelta = 0; /** * Create an aligner object. The aligner object will load and hold the BWT until close() is called. */ @Override public void initialize() { BWACConfiguration configuration = new BWACConfiguration(prefix); aligner = new BWACAligner(configuration); } /** Must return true for reads that need to be processed. Reads, for which this method return false will * be skipped by the engine and never passed to the walker. */ @Override public boolean filter(char[] ref, SAMRecord read) { return true; //return read.getReadName().contains("SL-XBC:1:61:1719:1512#0"); } /** * Aligns a read to the given reference. * @param ref Reference over the read. Read will most likely be unmapped, so ref will be null. * @param read Read to align. * @return Number of reads aligned by this map (aka 1). */ @Override public Integer map(char[] ref, SAMRecord read) { //logger.info(String.format("examining read %s", read.getReadName())); byte[] bases = read.getReadBases(); if(read.getReadNegativeStrandFlag()) bases = BaseUtils.simpleReverseComplement(bases); boolean matches = true; Alignment[] alignments = aligner.getAlignments(bases); if(alignments.length == 0 && !read.getReadUnmappedFlag()) matches = false; else { for(Alignment alignment: alignments) { matches = (alignment.getContigIndex() == read.getReferenceIndex()); matches &= (alignment.getAlignmentStart() == read.getAlignmentStart()); matches &= (alignment.isNegativeStrand() == read.getReadNegativeStrandFlag()); matches &= (alignment.getCigar().equals(read.getCigar())); int mapQDelta = Math.abs(alignment.getMappingQuality()-read.getMappingQuality()); maxMapQDelta = Math.max(mapQDelta,maxMapQDelta); matches &= (alignment.getMappingQuality() == read.getMappingQuality()); if(matches) break; } } if(!matches) { logger.error("Found mismatch!"); logger.error(String.format("Read %s:",read.getReadName())); logger.error(String.format(" Contig index: %d",read.getReferenceIndex())); logger.error(String.format(" Alignment start: %d", read.getAlignmentStart())); logger.error(String.format(" Negative strand: %b", read.getReadNegativeStrandFlag())); logger.error(String.format(" Cigar: %s%n", read.getCigarString())); logger.error(String.format(" Mapping quality: %s%n", read.getMappingQuality())); for(int i = 0; i < alignments.length; i++) { logger.error(String.format("Alignment %d:",i)); logger.error(String.format(" Contig index: %d",alignments[i].getContigIndex())); logger.error(String.format(" Alignment start: %d", alignments[i].getAlignmentStart())); logger.error(String.format(" Negative strand: %b", alignments[i].isNegativeStrand())); logger.error(String.format(" Cigar: %s", alignments[i].getCigarString())); logger.error(String.format(" Mapping quality: %s%n", alignments[i].getMappingQuality())); } throw new StingException(String.format("Read %s mismatches!", read.getReadName())); } if(++count % 10000 == 0) logger.info(String.format("Processed %d reads", count)); //logger.info(String.format("validated read %s", read.getReadName())); return 1; } /** * Initial value for reduce. In this case, validated reads will be counted. * @return 0, indicating no reads yet validated. */ @Override public Integer reduceInit() { return 0; } /** * Calculates the number of reads processed. * @param value Number of reads processed by this map. * @param sum Number of reads processed before this map. * @return Number of reads processed up to and including this map. */ @Override public Integer reduce(Integer value, Integer sum) { return value + sum; } /** * Cleanup. * @param result Number of reads processed. */ @Override public void onTraversalDone(Integer result) { aligner.close(); out.printf("Max mapping quality delta: %s%n", maxMapQDelta); super.onTraversalDone(result); } }