Пример #1
0
  private static void resetAttributes(
      SAMRecord rec, List<String> optFieldTags, List<Object> optFieldValues) {
    ListIterator<String> iterTags = optFieldTags.listIterator();
    ListIterator<Object> iterValues = optFieldValues.listIterator();

    while (iterTags.hasNext()) {
      rec.setAttribute(iterTags.next(), iterValues.next());
    }
  }
Пример #2
0
  private static void updateSAM(
      SAMRecord rec,
      ReferenceSequence sequence,
      SAMProgramRecord programRecord,
      AlignHeapNode bestAlignHeapNode,
      SRMAUtil.Space space,
      String read,
      String qualities,
      String softClipStartBases,
      String softClipStartQualities,
      String softClipEndBases,
      String softClipEndQualities,
      boolean strand,
      boolean correctBases)
      throws Exception {
    AlignHeapNode curAlignHeapNode = null;
    AlignHeapNode prevAlignHeapNode = null;

    int alignmentStart = 0;
    int readIndex = -1;
    byte readBases[] = null;
    byte baseQualities[] = null;
    byte colorErrors[] = null;
    int i;
    int numEdits = 0;
    List<String> optFieldTags = new LinkedList<String>();
    List<Object> optFieldValues = new LinkedList<Object>();
    Object attr;

    // Debugging stuff
    String readName = rec.getReadName();

    if (null == bestAlignHeapNode) {
      // Do not modify the alignment
      return;
    }

    // To generate a new CIGAR
    List<CigarElement> cigarElements = null;
    CigarOperator prevCigarOperator = null, curCigarOperator = null;
    int prevCigarOperatorLength = 0;

    // TODO
    // setInferredInsertSize (invalidates paired end reads)
    // setMappingQuality (?)
    // setFlag
    // update base qualities for color space reads

    // clear attributes, but save some
    Align.clearAttributes(rec, optFieldTags, optFieldValues);

    readBases = new byte[read.length()];
    baseQualities = new byte[qualities.length()];
    for (i = 0; i < qualities.length(); i++) {
      // Must subtract 33 for PHRED scaling
      baseQualities[i] = (byte) (qualities.charAt(i) - 33);
    }

    if (strand) {
      readIndex = 0;
    } else {
      readIndex = read.length() - 1;
    }
    cigarElements = new LinkedList<CigarElement>();
    if (strand) { // reverse strand is the current position
      alignmentStart = bestAlignHeapNode.node.position;
    } else {
      alignmentStart = bestAlignHeapNode.startPosition;
    }

    assert null != bestAlignHeapNode;
    curAlignHeapNode = bestAlignHeapNode;

    while (null != curAlignHeapNode) {
      // Get the current cigar operator
      if (null != prevAlignHeapNode
          && CigarOperator.DELETION != prevCigarOperator
          && 1 < Math.abs(curAlignHeapNode.node.position - prevAlignHeapNode.node.position)) {
        curCigarOperator = CigarOperator.DELETION;
      } else {
        switch (curAlignHeapNode.node.type) {
          case Node.MISMATCH: // Fall through
          case Node.MATCH:
            curCigarOperator = CigarOperator.MATCH_OR_MISMATCH;
            break;
          case Node.INSERTION:
            // System.out.println("INS");
            curCigarOperator = CigarOperator.INSERTION;
            break;
          default:
            throw new Exception("Unknown node type");
        }
        if (space == SRMAUtil.Space.COLORSPACE || correctBases) {
          readBases[readIndex] = (byte) curAlignHeapNode.node.base;
          if (strand) {
            readIndex++;
          } else {
            readIndex--;
          }
          // count the number of mismatches
          switch (curAlignHeapNode.node.type) {
            case Node.MISMATCH:
            case Node.INSERTION:
              numEdits++;
              break;
            default:
              break;
          }
        } else {
          // count the number of mismatches
          switch (curAlignHeapNode.node.type) {
            case Node.MATCH:
              if (read.charAt(curAlignHeapNode.readOffset) != curAlignHeapNode.node.base) {
                numEdits++;
              }
              break;
            case Node.MISMATCH: // Fall through
              if (read.charAt(curAlignHeapNode.readOffset)
                  != sequence.getBases()[curAlignHeapNode.node.position - 1]) {
                numEdits++;
              }
              break;
            case Node.INSERTION:
              numEdits++;
              break;
            default:
              break;
          }
        }
      }
      if (prevCigarOperator != curCigarOperator) {
        // different cigar operator

        // add the previous cigar operator
        if (null != prevCigarOperator) {
          if (strand) { // reverse
            // append
            cigarElements.add(new CigarElement(prevCigarOperatorLength, prevCigarOperator));
          } else {
            // prepend
            cigarElements.add(0, new CigarElement(prevCigarOperatorLength, prevCigarOperator));
          }
        }

        // update prevCigarOperator
        prevCigarOperator = curCigarOperator;
        if (curCigarOperator == CigarOperator.DELETION) {
          // length of deletion
          prevCigarOperatorLength =
              Math.abs(curAlignHeapNode.node.position - prevAlignHeapNode.node.position) - 1;
          numEdits += prevCigarOperatorLength; // deletions
        } else {
          prevCigarOperatorLength = 1;
        }
      } else {
        // same cigar operator
        prevCigarOperatorLength++;
      }

      // Update
      if (CigarOperator.DELETION != curCigarOperator) {
        prevAlignHeapNode = curAlignHeapNode;
        curAlignHeapNode = curAlignHeapNode.prev;
      }
    }
    if (0 < prevCigarOperatorLength) {
      if (null == prevCigarOperator || CigarOperator.DELETION == prevCigarOperator) {
        throw new Exception("Ended with a null cigar operator or a deletion cigar operator");
      }
      if (strand) { // reverse
        // append
        cigarElements.add(new CigarElement(prevCigarOperatorLength, prevCigarOperator));
      } else {
        // prepend
        cigarElements.add(0, new CigarElement(prevCigarOperatorLength, prevCigarOperator));
      }
    }

    if (space == SRMAUtil.Space.COLORSPACE) { // color space, read bases already inferred
      // Get color error string
      colorErrors = new byte[read.length()];
      char prevBase = SRMAUtil.COLORSPACE_ADAPTOR;
      if (strand) { // reverse
        for (i = 0; i < read.length(); i++) {
          char nextBase = SRMAUtil.colorSpaceNextBase(prevBase, read.charAt(i));
          if (nextBase == SRMAUtil.getCompliment((char) readBases[read.length() - i - 1])) {
            colorErrors[i] = (byte) Alignment.GAP;
          } else {
            colorErrors[i] = (byte) read.charAt(i);
          }
          if (0 < i) {
            // qualities are assumed to be always in the same direction as the color errors
            baseQualities[read.length() - i] =
                getColorQuality(
                    colorErrors[i - 1],
                    colorErrors[i],
                    (byte) (qualities.charAt(i - 1) - 33),
                    (byte) (qualities.charAt(i) - 33));
          }
          prevBase = SRMAUtil.getCompliment((char) readBases[read.length() - i - 1]);
        }
        // last color
        baseQualities[0] = (byte) (qualities.charAt(read.length() - 1) - 33);
      } else {
        for (i = 0; i < read.length(); i++) {
          char nextBase = SRMAUtil.colorSpaceNextBase(prevBase, read.charAt(i));
          if (nextBase == readBases[i]) {
            colorErrors[i] = (byte) Alignment.GAP;
          } else {
            colorErrors[i] = (byte) read.charAt(i);
          }
          if (0 < i) {
            baseQualities[i - 1] =
                getColorQuality(
                    colorErrors[i - 1],
                    colorErrors[i],
                    (byte) (qualities.charAt(i - 1) - 33),
                    (byte) (qualities.charAt(i) - 33));
          }
          prevBase = (char) readBases[i];
        }
        // last color
        baseQualities[read.length() - 1] = (byte) (qualities.charAt(read.length() - 1) - 33);
      }
    } else if (correctBases) { // bases were corrected
      if (strand) {
        for (i = 0; i < read.length(); i++) {
          if (readBases[i] == (byte) read.charAt(read.length() - i - 1)) {
            baseQualities[i] = (byte) (qualities.charAt(read.length() - i - 1) - 33);
          } else {
            // TODO: how much to down-weight ?
            baseQualities[i] =
                (byte)
                    (SRMAUtil.QUAL2CHAR(
                            SRMAUtil.CHAR2QUAL(qualities.charAt(read.length() - i - 1))
                                - CORRECT_BASE_QUALITY_PENALTY)
                        - 33);
            if (baseQualities[i] <= 0) {
              baseQualities[i] = 1;
            }
          }
        }
      } else {
        for (i = 0; i < read.length(); i++) {
          if (readBases[i] == (byte) read.charAt(i)) {
            baseQualities[i] = (byte) (qualities.charAt(i) - 33);
          } else {
            // TODO: how much to down-weight ?
            baseQualities[i] =
                (byte)
                    (SRMAUtil.QUAL2CHAR(
                            SRMAUtil.CHAR2QUAL(qualities.charAt(i)) - CORRECT_BASE_QUALITY_PENALTY)
                        - 33);
            if (baseQualities[i] <= 0) {
              baseQualities[i] = 1;
            }
          }
        }
      }
      rec.setAttribute("XO", read);
      rec.setAttribute("XQ", qualities);
    } else { // bases not corrected
      readBases = new byte[read.length()];
      baseQualities = new byte[qualities.length()]; // qualities.length() == read.length()
      if (strand) { // reverse
        for (i = 0; i < read.length(); i++) {
          readBases[i] = (byte) read.charAt(read.length() - i - 1);
          baseQualities[i] = (byte) (qualities.charAt(read.length() - i - 1) - 33);
        }
      } else {
        for (i = 0; i < read.length(); i++) {
          readBases[i] = (byte) read.charAt(i);
          baseQualities[i] = (byte) (qualities.charAt(i) - 33);
        }
      }
    }

    // Add in soft-clipping
    if (null != softClipStartBases) { // prepend
      cigarElements.add(0, new CigarElement(softClipStartBases.length(), CigarOperator.S));

      byte tmpBases[] = new byte[readBases.length + softClipStartBases.length()];
      System.arraycopy(readBases, 0, tmpBases, softClipStartBases.length(), readBases.length);
      readBases = tmpBases;
      for (i = 0; i < softClipStartBases.length(); i++) {
        readBases[i] = (byte) softClipStartBases.charAt(i);
      }

      byte tmpQualities[] = new byte[baseQualities.length + softClipStartQualities.length()];
      System.arraycopy(
          baseQualities, 0, tmpQualities, softClipStartQualities.length(), baseQualities.length);
      baseQualities = tmpQualities;
      for (i = 0; i < softClipStartQualities.length(); i++) {
        baseQualities[i] = (byte) softClipStartQualities.charAt(i);
      }
    }
    if (null != softClipEndBases) { // append
      cigarElements.add(new CigarElement(softClipEndBases.length(), CigarOperator.S));

      byte tmpBases[] = new byte[readBases.length + softClipEndBases.length()];
      System.arraycopy(readBases, 0, tmpBases, 0, readBases.length);
      for (i = 0; i < softClipEndBases.length(); i++) {
        tmpBases[i + readBases.length] = (byte) softClipEndBases.charAt(i);
      }
      readBases = tmpBases;

      byte tmpQualities[] = new byte[baseQualities.length + softClipEndQualities.length()];
      System.arraycopy(baseQualities, 0, tmpQualities, 0, baseQualities.length);
      for (i = 0; i < softClipEndQualities.length(); i++) {
        tmpQualities[i + baseQualities.length] = (byte) softClipEndQualities.charAt(i);
      }
      baseQualities = tmpQualities;
    }

    // Update SAM record
    rec.setCigar(new Cigar(cigarElements));
    rec.setAlignmentStart(alignmentStart);
    rec.setReadBases(readBases);
    rec.setBaseQualities(baseQualities);
    // Reset saved attributes
    Align.resetAttributes(rec, optFieldTags, optFieldValues);
    // Set new attributes
    if (space == SRMAUtil.Space.COLORSPACE) {
      // set the XE attribute for colorError string
      rec.setAttribute("XE", new String(colorErrors));
    }
    rec.setAttribute("AS", bestAlignHeapNode.score);
    rec.setAttribute("XC", bestAlignHeapNode.alleleCoverageSum);
    rec.setAttribute("PG", programRecord.getId());
    rec.setAttribute("NM", numEdits);
  }
  @Override
  public void execute() {
    log.info("Initializing kmer code map...");
    Map<Character, Integer> kmerCodeIndices = new HashMap<Character, Integer>();
    kmerCodeIndices.put('0', 1);
    kmerCodeIndices.put('A', 3);
    kmerCodeIndices.put('B', 4);
    kmerCodeIndices.put('C', 5);
    kmerCodeIndices.put('_', 6);
    kmerCodeIndices.put('.', 7);
    kmerCodeIndices.put('1', 9);

    Map<Character, String> kmerCodeNames = new LinkedHashMap<Character, String>();
    kmerCodeNames.put('0', "ref0");
    kmerCodeNames.put('A', "repetitive");
    kmerCodeNames.put('B', "both");
    kmerCodeNames.put('C', "lowcoverage");
    kmerCodeNames.put('_', "lowconfidence");
    kmerCodeNames.put('.', "novel");
    kmerCodeNames.put('1', "ref1");

    if (KMER_CODE_NAMES != null) {
      for (Character c : kmerCodeNames.keySet()) {
        String cStr = String.valueOf(c);
        if (KMER_CODE_NAMES.containsKey(cStr)) {
          kmerCodeNames.put(c, KMER_CODE_NAMES.get(cStr));
        }
      }
    }

    for (Character c : kmerCodeNames.keySet()) {
      log.info("  {} {}: {}", c, kmerCodeIndices.get(c), kmerCodeNames.get(c));
    }

    log.info("Loading annotated contigs...");
    Map<String, Map<String, String>> annotatedContigs = new HashMap<String, Map<String, String>>();
    int kmerSize = 0;

    if (ANN.length() > 0) {
      TableReader tr = new TableReader(ANN);
      for (Map<String, String> te : tr) {
        String contigName = te.get("contigName");

        if (kmerSize == 0) {
          kmerSize = te.get("seq").length() - te.get("kmerOrigin").length() + 1;
        }

        annotatedContigs.put(contigName, te);

        String[] ref0ToCanonicalExact =
            (te.get("ref0ToCanonicalExact").equals("NA")
                        || te.get("ref0ToCanonicalExact").equals("*:0-0")
                    ? "NA:0-0"
                    : te.get("ref0ToCanonicalExact"))
                .split("[:-]");
        String[] ref1ToCanonicalExact =
            (te.get("ref1ToCanonicalExact").equals("NA")
                        || te.get("ref1ToCanonicalExact").equals("*:0-0")
                    ? "NA:0-0"
                    : te.get("ref1ToCanonicalExact"))
                .split("[:-]");

        cout.println(
            te.get("sampleName")
                + "_"
                + te.get("accession")
                + "_"
                + contigName
                + " "
                + ref0ToCanonicalExact[0]
                + " "
                + ref0ToCanonicalExact[1]
                + " "
                + ref0ToCanonicalExact[2]
                + " radius1=0.8r");
        cout.println(
            te.get("sampleName")
                + "_"
                + te.get("accession")
                + "_"
                + contigName
                + " "
                + ref1ToCanonicalExact[0]
                + " "
                + ref1ToCanonicalExact[1]
                + " "
                + ref1ToCanonicalExact[2]
                + " radius2=0.6r");
      }
    }

    log.info("    contigs: {}", annotatedContigs.size());
    log.info("  kmer size: {}", kmerSize);

    log.info("Computing kmer inheritance information...");

    SAMFileHeader sfh = CONTIGS.getFileHeader();
    for (Character c : kmerCodeNames.keySet()) {
      SAMReadGroupRecord rgr = new SAMReadGroupRecord(kmerCodeNames.get(c));
      rgr.setSample(kmerCodeNames.get(c));
      sfh.addReadGroup(rgr);
    }

    SAMFileWriterFactory sfwf = new SAMFileWriterFactory();
    sfwf.setCreateIndex(true);
    SAMFileWriter sfw = sfwf.makeBAMWriter(sfh, false, bout);

    TableWriter tw = new TableWriter(sout);

    Set<IGVEntry> igvEntries = new TreeSet<IGVEntry>();
    int numContigs = 0;
    for (SAMRecord contig : CONTIGS) {
      if (CONTIG_NAMES == null
          || CONTIG_NAMES.isEmpty()
          || CONTIG_NAMES.contains(contig.getReadName())) {
        Map<String, String> te = annotatedContigs.get(contig.getReadName());

        if (annotatedContigs.containsKey(contig.getReadName())) {
          String seq = contig.getReadString();

          // log.debug("  te: {}", te);

          String annSeq = te.get("seq");
          String kmerOrigin = te.get("kmerOrigin");

          Map<CortexKmer, Character> kmerCodes = new HashMap<CortexKmer, Character>();
          for (int i = 0; i < kmerOrigin.length(); i++) {
            CortexKmer kmer = new CortexKmer(annSeq.substring(i, i + kmerSize));
            Character code = kmerOrigin.charAt(i);

            kmerCodes.put(kmer, code);
          }

          Map<Character, Integer> kmerStats = new HashMap<Character, Integer>();
          for (Character c : kmerCodeNames.keySet()) {
            kmerStats.put(c, 0);
          }

          boolean changed = false;

          // We want to be able to examine soft-clipped regions as well.
          List<CigarElement> ces = new ArrayList<CigarElement>();
          for (CigarElement ce : contig.getCigar().getCigarElements()) {
            if (ce.getOperator().equals(CigarOperator.S)) {
              ces.add(new CigarElement(ce.getLength(), CigarOperator.M));
              changed = true;
            } else {
              ces.add(ce);
            }
          }

          if (changed) {
            CigarElement firstCe = contig.getCigar().getCigarElements().get(0);

            if (firstCe.getOperator().equals(CigarOperator.S)) {
              contig.setAlignmentStart(contig.getAlignmentStart() - firstCe.getLength());
            }

            contig.setCigar(new Cigar(ces));
          }

          for (AlignmentBlock ab : contig.getAlignmentBlocks()) {
            for (int i = ab.getReadStart() - 1; i < ab.getReadStart() + ab.getLength(); i++) {
              if (i + kmerSize < seq.length()) {
                CortexKmer kmer = new CortexKmer(seq.substring(i, i + kmerSize));

                SAMRecord skmer = new SAMRecord(CONTIGS.getFileHeader());
                skmer.setReadBases(seq.substring(i, i + kmerSize).getBytes());

                List<CigarElement> cigarElements = new ArrayList<CigarElement>();
                cigarElements.add(new CigarElement(kmerSize, CigarOperator.M));
                Cigar cigar = new Cigar(cigarElements);

                skmer.setReadName(contig.getReadName() + "." + kmer.getKmerAsString());
                skmer.setReferenceName(contig.getReferenceName());
                skmer.setCigar(cigar);
                skmer.setReadPairedFlag(false);
                skmer.setDuplicateReadFlag(false);
                skmer.setMateNegativeStrandFlag(false);
                skmer.setAlignmentStart(ab.getReferenceStart() - ab.getReadStart() + 1 + i);
                skmer.setAttribute("RG", "none");
                skmer.setMappingQuality(0);

                Character c = kmerCodes.get(kmer);
                String codeName = kmerCodeNames.get(c);

                String parentReadGroupId = null;
                String sampleReadGroupId = null;
                for (SAMReadGroupRecord rgr : sfh.getReadGroups()) {
                  if (rgr.getSample().equals(codeName)) {
                    parentReadGroupId = rgr.getReadGroupId();
                  }

                  if (rgr.getSample().equals(contig.getReadGroup().getSample())) {
                    sampleReadGroupId = rgr.getReadGroupId();
                  }
                }

                skmer.setAttribute(
                    "RG", parentReadGroupId != null ? parentReadGroupId : sampleReadGroupId);
                skmer.setMappingQuality(99);

                sfw.addAlignment(skmer);

                kmerStats.put(c, kmerStats.get(c) + 1);

                IGVEntry igvEntry = new IGVEntry();
                igvEntry.chromosome = contig.getReferenceName();
                igvEntry.start = ab.getReferenceStart() - ab.getReadStart() + i;
                igvEntry.parentageName = kmerCodeNames.get(c);
                igvEntry.parentage = kmerCodeIndices.get(c);
                igvEntries.add(igvEntry);
              }
            }
          }

          if (!contig.isSecondaryOrSupplementary()) {
            beout.println(
                contig.getReferenceName()
                    + "\t"
                    + contig.getAlignmentStart()
                    + "\t"
                    + contig.getAlignmentEnd()
                    + "\t"
                    + contig.getReadName()
                    + "."
                    + contig.getReadGroup().getSample());

            if (annotatedContigs.size() > 10 && numContigs % (annotatedContigs.size() / 10) == 0) {
              log.info("  processed {}/{} contigs", numContigs, annotatedContigs.size());
            }
            numContigs++;
          }

          Map<String, String> stats = new LinkedHashMap<String, String>();
          stats.put("contigName", contig.getReadName());
          stats.put("sampleName", contig.getReadGroup().getSample());
          for (Character c : kmerCodeNames.keySet()) {
            stats.put(kmerCodeNames.get(c), String.valueOf(kmerStats.get(c)));
          }
          tw.addEntry(stats);
        }
      }
    }

    log.info("Writing kmer inheritance information...");
    out.printf("%s\t%s\t%s\t%s\t%s\n", "Chromosome", "Start", "End", "Feature", "Parentage");
    for (IGVEntry igvEntry : igvEntries) {
      out.printf(
          "%s\t%d\t%d\t%s\t%d\n",
          igvEntry.chromosome,
          igvEntry.start,
          igvEntry.start + 1,
          igvEntry.parentageName,
          igvEntry.parentage);
    }

    sfw.close();
  }