package org.broadinstitute.sting.utils; import net.sf.samtools.CigarOperator; import net.sf.samtools.SAMRecord; import net.sf.samtools.Cigar; import net.sf.samtools.CigarElement; import net.sf.picard.reference.ReferenceSequence; import org.broadinstitute.sting.playground.utils.CountedObject; import java.util.*; /** * Created by IntelliJ IDEA. * User: asivache * Date: Mar 25, 2009 * Time: 12:15:38 AM * To change this template use File | Settings | File Templates. */ public class AlignmentUtils { /** Returns number of mismatches in the alignment r to the reference sequence * refSeq. It is assumed that * the alignment starts at (1-based) position r.getAlignmentStart() on the specified, and all single-base mismatches * are counted in the alignment segments where both sequences are present. Insertions/deletions are skipped and do * not contribute to the error count returned by this method. * @param r aligned read * @param refSeq reference sequence * @return number of single-base mismatches in the aligned segments (gaps on either of the sequences are skipped) */ public static int numMismatches(SAMRecord r, ReferenceSequence refSeq) { byte[] ref = refSeq.getBases(); if ( r.getReadUnmappedFlag() ) return 1000000; int i_ref = r.getAlignmentStart()-1; // position on the ref int i_read = 0; // position on the read int mm_count = 0; // number of mismatches Cigar c = r.getCigar(); for ( int k = 0 ; k < c.numCigarElements() ; k++ ) { CigarElement ce = c.getCigarElement(k); switch( ce.getOperator() ) { case M: for ( int l = 0 ; l < ce.getLength() ; l++, i_ref++, i_read++ ) { char refChr = (char)ref[i_ref]; char readChr = r.getReadString().charAt(i_read); if ( BaseUtils.simpleBaseToBaseIndex(readChr) == -1 || BaseUtils.simpleBaseToBaseIndex(refChr) == -1 ) continue; // do not count Ns/Xs/etc ? if ( Character.toUpperCase(readChr) != Character.toUpperCase(refChr) ) mm_count++; } break; case I: i_read += ce.getLength(); break; case D: i_ref += ce.getLength(); break; default: throw new RuntimeException("Unrecognized cigar element"); } } return mm_count; } /** * mhanna - 11 May 2009 - stubbed out competing method that works with partial references. * Computes number of mismatches in the read alignment to the refence ref * specified in the record r. Indels are completely ignored by this method: * only base mismatches in the alignment segments where both sequences are present are counted. * @param r * @return */ public static int numMismatches(SAMRecord r, char[] ref) { if ( r.getReadUnmappedFlag() ) return 1000000; int i_ref = 0; // position on the ref int i_read = 0; // position on the read int mm_count = 0; // number of mismatches Cigar c = r.getCigar(); for ( int k = 0 ; k < c.numCigarElements() ; k++ ) { CigarElement ce = c.getCigarElement(k); switch( ce.getOperator() ) { case M: for ( int l = 0 ; l < ce.getLength() ; l++, i_ref++, i_read++ ) { char refChr = (char)ref[i_ref]; char readChr = r.getReadString().charAt(i_read); if ( BaseUtils.simpleBaseToBaseIndex(readChr) == -1 || BaseUtils.simpleBaseToBaseIndex(refChr) == -1 ) continue; // do not count Ns/Xs/etc ? if ( Character.toUpperCase(readChr) != Character.toUpperCase(refChr) ) mm_count++; } break; case I: i_read += ce.getLength(); break; case D: i_ref += ce.getLength(); break; default: throw new RuntimeException("Unrecognized cigar element"); } } return mm_count; } // IMPORTANT NOTE: ALTHOUGH THIS METHOD IS EXTREMELY SIMILAR TO THE ONE ABOVE, WE NEED // TWO SEPARATE IMPLEMENTATIONS IN ORDER TO PREVENT JAVA STRINGS FROM FORCING US TO // PERFORM EXPENSIVE ARRAY COPYING WHEN TRYING TO GET A BYTE ARRAY... /** See {@link #numMismatches(SAMRecord, ReferenceSequence)}. This method implements same functionality * for reference sequence specified as conventional java string (of bases). By default, it is assumed that * the alignment starts at (1-based) position r.getAlignmentStart() on the reference refSeq. * See {@link #numMismatches(SAMRecord, String, int)} if this is not the case. */ public static int numMismatches(SAMRecord r, String refSeq ) { if ( r.getReadUnmappedFlag() ) return 1000000; return numMismatches(r, refSeq, r.getAlignmentStart()-1); } /** Returns number of mismatches in the alignment r to the reference sequence * refSeq assuming the alignment starts at (ZERO-based) position refIndex on the * specified reference sequence; in other words, refIndex is used in place of alignment's own * getAlignmentStart() coordinate and the latter is never used. However, the structure of the alignment r * (i.e. it's cigar string with all the insertions/deletions it may specify) is fully respected. * * @param r alignment * @param refSeq chunk of reference sequence that subsumes the alignment completely (if alignment runs out of * the reference string, IndexOutOfBound exception will be thrown at runtime). * @param refIndex zero-based position, at which the alignment starts on the specified reference string. * @return */ public static int numMismatches(SAMRecord r, String refSeq, int refIndex) { int readIdx = 0; int mismatches = 0; String readSeq = r.getReadString(); Cigar c = r.getCigar(); for (int i = 0 ; i < c.numCigarElements() ; i++) { CigarElement ce = c.getCigarElement(i); switch ( ce.getOperator() ) { case M: for (int j = 0 ; j < ce.getLength() ; j++, refIndex++, readIdx++ ) { if ( refIndex >= refSeq.length() ) continue; char refChr = refSeq.charAt(refIndex); char readChr = readSeq.charAt(readIdx); // Note: we need to count X/N's as mismatches because that's what SAM requires //if ( BaseUtils.simpleBaseToBaseIndex(readChr) == -1 || // BaseUtils.simpleBaseToBaseIndex(refChr) == -1 ) // continue; // do not count Ns/Xs/etc ? if ( Character.toUpperCase(readChr) != Character.toUpperCase(refChr) ) mismatches++; } break; case I: readIdx += ce.getLength(); break; case D: refIndex += ce.getLength(); break; default: throw new StingException("Only M,I,D cigar elements are currently supported"); } } return mismatches; } /** Returns number of alignment blocks (continuous stretches of aligned bases) in the specified alignment. * This method follows closely the SAMRecord::getAlignmentBlocks() implemented in samtools library, but * it only counts blocks without actually allocating and filling the list of blocks themselves. Hence, this method is * a much more efficient alternative to r.getAlignmentBlocks.size() in the situations when this number is all that is needed. * Formally, this method simply returns the number of M elements in the cigar. * @param r alignment * @return number of continuous alignment blocks (i.e. 'M' elements of the cigar; all indel and clipping elements are ignored). */ public static int getNumAlignmentBlocks(final SAMRecord r) { int n = 0; final Cigar cigar = r.getCigar(); if (cigar == null) return 0; for (final CigarElement e : cigar.getCigarElements()) { if (e.getOperator() == CigarOperator.M ) n++; } return n; } public static String toString(Cigar cig) { StringBuilder b = new StringBuilder(); for ( int i = 0 ; i < cig.numCigarElements() ; i++ ) { char c='?'; switch ( cig.getCigarElement(i).getOperator() ) { case M : c = 'M'; break; case D : c = 'D'; break; case I : c = 'I'; break; } b.append(cig.getCigarElement(i).getLength()); b.append(c); } return b.toString(); } public static String alignmentToString(final Cigar cigar,final String seq, final String ref, final int posOnRef ) { return alignmentToString( cigar, seq, ref, posOnRef, 0 ); } public static String cigarToString(Cigar cig) { if ( cig == null ) return "null"; StringBuilder b = new StringBuilder(); for ( int i = 0 ; i < cig.numCigarElements() ; i++ ) { char c='?'; switch ( cig.getCigarElement(i).getOperator() ) { case M : c = 'M'; break; case D : c = 'D'; break; case I : c = 'I'; break; } b.append(cig.getCigarElement(i).getLength()); b.append(c); } return b.toString(); } public static String alignmentToString(final Cigar cigar,final String seq, final String ref, final int posOnRef, final int posOnRead ) { int readPos = posOnRead; int refPos = posOnRef; StringBuilder refLine = new StringBuilder(); StringBuilder readLine = new StringBuilder(); for ( int i = 0 ; i < posOnRead ; i++ ) { refLine.append( ref.charAt( refPos - readPos + i ) ); readLine.append( seq.charAt(i) ) ; } for ( int i = 0 ; i < cigar.numCigarElements() ; i++ ) { final CigarElement ce = cigar.getCigarElement(i); switch(ce.getOperator()) { case I: for ( int j = 0 ; j < ce.getLength(); j++ ) { refLine.append('+'); readLine.append( seq.charAt( readPos++ ) ); } break; case D: for ( int j = 0 ; j < ce.getLength(); j++ ) { readLine.append('*'); refLine.append( ref.charAt( refPos++ ) ); } break; case M: for ( int j = 0 ; j < ce.getLength(); j++ ) { refLine.append(ref.charAt( refPos++ ) ); readLine.append( seq.charAt( readPos++ ) ); } break; default: throw new StingException("Unsupported cigar operator: "+ce.getOperator() ); } } refLine.append('\n'); refLine.append(readLine); refLine.append('\n'); return refLine.toString(); } }