public static boolean allelesAreSubset(VariantContext vc1, VariantContext vc2) {
// if all alleles of vc1 are a contained in alleles of vc2, return true
if (!vc1.getReference().equals(vc2.getReference())) return false;
for (Allele a : vc1.getAlternateAlleles()) {
if (!vc2.getAlternateAlleles().contains(a)) return false;
}
return true;
}
/**
* Update the attributes of the attributes map given the VariantContext to reflect the proper
* chromosome-based VCF tags
*
* @param vc the VariantContext
* @param attributes the attributes map to populate; must not be null; may contain old values
* @param removeStaleValues should we remove stale values from the mapping?
* @return the attributes map provided as input, returned for programming convenience
*/
public static Map<String, Object> calculateChromosomeCounts(
VariantContext vc, Map<String, Object> attributes, boolean removeStaleValues) {
final int AN = vc.getCalledChrCount();
// if everyone is a no-call, remove the old attributes if requested
if (AN == 0 && removeStaleValues) {
if (attributes.containsKey(VCFConstants.ALLELE_COUNT_KEY))
attributes.remove(VCFConstants.ALLELE_COUNT_KEY);
if (attributes.containsKey(VCFConstants.ALLELE_FREQUENCY_KEY))
attributes.remove(VCFConstants.ALLELE_FREQUENCY_KEY);
if (attributes.containsKey(VCFConstants.ALLELE_NUMBER_KEY))
attributes.remove(VCFConstants.ALLELE_NUMBER_KEY);
return attributes;
}
if (vc.hasGenotypes()) {
attributes.put(VCFConstants.ALLELE_NUMBER_KEY, AN);
// if there are alternate alleles, record the relevant tags
if (vc.getAlternateAlleles().size() > 0) {
final ArrayList<String> alleleFreqs = new ArrayList<String>();
final ArrayList<Integer> alleleCounts = new ArrayList<Integer>();
for (Allele allele : vc.getAlternateAlleles()) {
int altChromosomes = vc.getCalledChrCount(allele);
alleleCounts.add(altChromosomes);
if (AN == 0) {
alleleFreqs.add("0.0");
} else {
// todo -- this is a performance problem
final String freq =
String.format(
makePrecisionFormatStringFromDenominatorValue((double) AN),
((double) altChromosomes / (double) AN));
alleleFreqs.add(freq);
}
}
attributes.put(
VCFConstants.ALLELE_COUNT_KEY,
alleleCounts.size() == 1 ? alleleCounts.get(0) : alleleCounts);
attributes.put(
VCFConstants.ALLELE_FREQUENCY_KEY,
alleleFreqs.size() == 1 ? alleleFreqs.get(0) : alleleFreqs);
} else {
attributes.put(VCFConstants.ALLELE_COUNT_KEY, 0);
attributes.put(VCFConstants.ALLELE_FREQUENCY_KEY, 0.0);
}
}
return attributes;
}
@Requires({"eval != null", "comp != null"})
private EvalCompMatchType doEvalAndCompMatch(
final VariantContext eval, final VariantContext comp, boolean requireStrictAlleleMatch) {
// find all of the matching comps
if (comp.getType() != eval.getType()) return EvalCompMatchType.NO_MATCH;
// find the comp which matches both the reference allele and alternate allele from eval
final Allele altEval =
eval.getAlternateAlleles().size() == 0 ? null : eval.getAlternateAllele(0);
final Allele altComp =
comp.getAlternateAlleles().size() == 0 ? null : comp.getAlternateAllele(0);
if ((altEval == null && altComp == null)
|| (altEval != null
&& altEval.equals(altComp)
&& eval.getReference().equals(comp.getReference()))) return EvalCompMatchType.STRICT;
else return requireStrictAlleleMatch ? EvalCompMatchType.NO_MATCH : EvalCompMatchType.LENIENT;
}
private boolean haveDifferentAltAlleles(VariantContext eval, VariantContext comp) {
Collection<Allele> evalAlts = eval.getAlternateAlleles();
Collection<Allele> compAlts = comp.getAlternateAlleles();
if (evalAlts.size() != compAlts.size()) {
return true;
} else {
// same size => every alt from eval must be in comp
for (Allele a : evalAlts) {
if (!compAlts.contains(a)) {
// System.out.printf("Different alleles: %s:%d eval=%s
// comp=%s\n\t\teval=%s\n\t\tcomp=%s%n",
// eval.getChr(), eval.getStart(), eval.getAlleles(),
// comp.getAlleles(), eval, comp);
return true;
}
}
return false;
}
}
private Map<String, Object> annotateSNP(AlignmentContext stratifiedContext, VariantContext vc) {
if (!stratifiedContext.hasBasePileup()) return null;
HashMap<Byte, Integer> alleleCounts = new HashMap<Byte, Integer>();
for (Allele allele : vc.getAlternateAlleles()) alleleCounts.put(allele.getBases()[0], 0);
ReadBackedPileup pileup = stratifiedContext.getBasePileup();
int totalDepth = pileup.size();
Map<String, Object> map = new HashMap<String, Object>();
map.put(getKeyNames().get(0), totalDepth); // put total depth in right away
if (totalDepth == 0) return map; // done, can not compute FA at 0 coverage!!
int mq0 = 0; // number of "ref" reads that are acually mq0
for (PileupElement p : pileup) {
if (p.getMappingQual() == 0) {
mq0++;
continue;
}
if (alleleCounts.containsKey(p.getBase())) // non-mq0 read and it's an alt
alleleCounts.put(p.getBase(), alleleCounts.get(p.getBase()) + 1);
}
if (mq0 == totalDepth) return map; // if all reads are mq0, there is nothing left to do
// we need to add counts in the correct order
String[] fracs = new String[alleleCounts.size()];
for (int i = 0; i < vc.getAlternateAlleles().size(); i++) {
fracs[i] =
String.format(
"%.3f",
((float) alleleCounts.get(vc.getAlternateAllele(i).getBases()[0]))
/ (totalDepth - mq0));
}
map.put(getKeyNames().get(1), fracs);
return map;
}
/**
* @param other VariantContext whose alternate alleles to compare against
* @return true if this VariantContext has the same alternate alleles as other, regardless of
* ordering. Otherwise returns false.
*/
public boolean hasSameAlternateAllelesAs(VariantContext other) {
List<Allele> thisAlternateAlleles = getAlternateAlleles();
List<Allele> otherAlternateAlleles = other.getAlternateAlleles();
if (thisAlternateAlleles.size() != otherAlternateAlleles.size()) {
return false;
}
for (Allele allele : thisAlternateAlleles) {
if (!otherAlternateAlleles.contains(allele)) {
return false;
}
}
return true;
}
public String update1(
VariantContext vc1,
RefMetaDataTracker tracker,
ReferenceContext ref,
AlignmentContext context) {
nCalledLoci++;
// Note from Eric:
// This is really not correct. What we really want here is a polymorphic vs. monomorphic count
// (i.e. on the Genotypes).
// So in order to maintain consistency with the previous implementation (and the intention of
// the original author), I've
// added in a proxy check for monomorphic status here.
// Protect against case when vc only as no-calls too - can happen if we strafity by sample and
// sample as a single no-call.
if (vc1.isMonomorphicInSamples()) {
nRefLoci++;
} else {
switch (vc1.getType()) {
case NO_VARIATION:
// shouldn't get here
break;
case SNP:
nVariantLoci++;
nSNPs++;
if (vc1.getAttributeAsBoolean("ISSINGLETON", false)) nSingletons++;
break;
case MNP:
nVariantLoci++;
nMNPs++;
if (vc1.getAttributeAsBoolean("ISSINGLETON", false)) nSingletons++;
break;
case INDEL:
nVariantLoci++;
if (vc1.isSimpleInsertion()) nInsertions++;
else if (vc1.isSimpleDeletion()) nDeletions++;
else nComplex++;
break;
case MIXED:
nVariantLoci++;
nMixed++;
break;
case SYMBOLIC:
nSymbolic++;
break;
default:
throw new ReviewedStingException("Unexpected VariantContext type " + vc1.getType());
}
}
String refStr = vc1.getReference().getBaseString().toUpperCase();
String aaStr =
vc1.hasAttribute("ANCESTRALALLELE")
? vc1.getAttributeAsString("ANCESTRALALLELE", null).toUpperCase()
: null;
// if (aaStr.equals(".")) {
// aaStr = refStr;
// }
// ref aa alt class
// A C A der homozygote
// A C C anc homozygote
// A A A ref homozygote
// A A C
// A C A
// A C C
for (final Genotype g : vc1.getGenotypes()) {
final String altStr =
vc1.getAlternateAlleles().size() > 0
? vc1.getAlternateAllele(0).getBaseString().toUpperCase()
: null;
switch (g.getType()) {
case NO_CALL:
nNoCalls++;
break;
case HOM_REF:
nHomRef++;
if (aaStr != null && altStr != null && !refStr.equalsIgnoreCase(aaStr)) {
nHomDerived++;
}
break;
case HET:
nHets++;
break;
case HOM_VAR:
nHomVar++;
if (aaStr != null && altStr != null && !altStr.equalsIgnoreCase(aaStr)) {
nHomDerived++;
}
break;
case MIXED:
break;
default:
throw new ReviewedStingException("BUG: Unexpected genotype type: " + g);
}
}
return null; // we don't capture any interesting sites
}
/**
* add the genotype data
*
* @param vc the variant context
* @param genotypeFormatKeys Genotype formatting string
* @param alleleMap alleles for this context
* @throws IOException for writer
*/
private void addGenotypeData(
VariantContext vc, Map<Allele, String> alleleMap, List<String> genotypeFormatKeys)
throws IOException {
for (String sample : mHeader.getGenotypeSamples()) {
mWriter.write(VCFConstants.FIELD_SEPARATOR);
Genotype g = vc.getGenotype(sample);
if (g == null) {
// TODO -- The VariantContext needs to know what the general ploidy is of the samples
// TODO -- We shouldn't be assuming diploid genotypes here!
mWriter.write(VCFConstants.EMPTY_GENOTYPE);
continue;
}
List<String> attrs = new ArrayList<String>(genotypeFormatKeys.size());
for (String key : genotypeFormatKeys) {
if (key.equals(VCFConstants.GENOTYPE_KEY)) {
if (!g.isAvailable()) {
throw new ReviewedStingException(
"GTs cannot be missing for some samples if they are available for others in the record");
}
writeAllele(g.getAllele(0), alleleMap);
for (int i = 1; i < g.getPloidy(); i++) {
mWriter.write(g.isPhased() ? VCFConstants.PHASED : VCFConstants.UNPHASED);
writeAllele(g.getAllele(i), alleleMap);
}
continue;
}
Object val = g.hasAttribute(key) ? g.getAttribute(key) : VCFConstants.MISSING_VALUE_v4;
// some exceptions
if (key.equals(VCFConstants.GENOTYPE_QUALITY_KEY)) {
if (Math.abs(g.getNegLog10PError() - Genotype.NO_NEG_LOG_10PERROR) < 1e-6)
val = VCFConstants.MISSING_VALUE_v4;
else {
val = getQualValue(Math.min(g.getPhredScaledQual(), VCFConstants.MAX_GENOTYPE_QUAL));
}
} else if (key.equals(VCFConstants.GENOTYPE_FILTER_KEY)) {
val =
g.isFiltered()
? ParsingUtils.join(";", ParsingUtils.sortList(g.getFilters()))
: (g.filtersWereApplied()
? VCFConstants.PASSES_FILTERS_v4
: VCFConstants.UNFILTERED);
}
VCFFormatHeaderLine metaData = mHeader.getFormatHeaderLine(key);
if (metaData != null) {
int numInFormatField = metaData.getCount(vc.getAlternateAlleles().size());
if (numInFormatField > 1 && val.equals(VCFConstants.MISSING_VALUE_v4)) {
// If we have a missing field but multiple values are expected, we need to construct a
// new string with all fields.
// For example, if Number=2, the string has to be ".,."
StringBuilder sb = new StringBuilder(VCFConstants.MISSING_VALUE_v4);
for (int i = 1; i < numInFormatField; i++) {
sb.append(",");
sb.append(VCFConstants.MISSING_VALUE_v4);
}
val = sb.toString();
}
}
// assume that if key is absent, then the given string encoding suffices
String outputValue = formatVCFField(val);
if (outputValue != null) attrs.add(outputValue);
}
// strip off trailing missing values
for (int i = attrs.size() - 1; i >= 0; i--) {
if (isMissingValue(attrs.get(i))) attrs.remove(i);
else break;
}
for (int i = 0; i < attrs.size(); i++) {
if (i > 0 || genotypeFormatKeys.contains(VCFConstants.GENOTYPE_KEY))
mWriter.write(VCFConstants.GENOTYPE_FIELD_SEPARATOR);
mWriter.write(attrs.get(i));
}
}
}
/**
* add a record to the file
*
* @param vc the Variant Context object
* @param refBase the ref base used for indels
* @param refBaseShouldBeAppliedToEndOfAlleles *** THIS SHOULD BE FALSE EXCEPT FOR AN INDEL AT THE
* EXTREME BEGINNING OF A CONTIG (WHERE THERE IS NO PREVIOUS BASE, SO WE USE THE BASE AFTER
* THE EVENT INSTEAD)
*/
public void add(VariantContext vc, byte refBase, boolean refBaseShouldBeAppliedToEndOfAlleles) {
if (mHeader == null)
throw new IllegalStateException(
"The VCF Header must be written before records can be added: " + locationString());
if (doNotWriteGenotypes) vc = VariantContext.modifyGenotypes(vc, null);
try {
vc =
VariantContext.createVariantContextWithPaddedAlleles(
vc, refBase, refBaseShouldBeAppliedToEndOfAlleles);
// if we are doing on the fly indexing, add the record ***before*** we write any bytes
if (indexer != null) indexer.addFeature(vc, positionalStream.getPosition());
Map<Allele, String> alleleMap = new HashMap<Allele, String>(vc.getAlleles().size());
alleleMap.put(Allele.NO_CALL, VCFConstants.EMPTY_ALLELE); // convenience for lookup
// CHROM
mWriter.write(vc.getChr());
mWriter.write(VCFConstants.FIELD_SEPARATOR);
// POS
mWriter.write(String.valueOf(vc.getStart()));
mWriter.write(VCFConstants.FIELD_SEPARATOR);
// ID
String ID = vc.hasID() ? vc.getID() : VCFConstants.EMPTY_ID_FIELD;
mWriter.write(ID);
mWriter.write(VCFConstants.FIELD_SEPARATOR);
// REF
alleleMap.put(vc.getReference(), "0");
String refString = vc.getReference().getDisplayString();
mWriter.write(refString);
mWriter.write(VCFConstants.FIELD_SEPARATOR);
// ALT
if (vc.isVariant()) {
Allele altAllele = vc.getAlternateAllele(0);
alleleMap.put(altAllele, "1");
String alt = altAllele.getDisplayString();
mWriter.write(alt);
for (int i = 1; i < vc.getAlternateAlleles().size(); i++) {
altAllele = vc.getAlternateAllele(i);
alleleMap.put(altAllele, String.valueOf(i + 1));
alt = altAllele.getDisplayString();
mWriter.write(",");
mWriter.write(alt);
}
} else {
mWriter.write(VCFConstants.EMPTY_ALTERNATE_ALLELE_FIELD);
}
mWriter.write(VCFConstants.FIELD_SEPARATOR);
// QUAL
if (!vc.hasNegLog10PError()) mWriter.write(VCFConstants.MISSING_VALUE_v4);
else mWriter.write(getQualValue(vc.getPhredScaledQual()));
mWriter.write(VCFConstants.FIELD_SEPARATOR);
// FILTER
String filters =
vc.isFiltered()
? ParsingUtils.join(";", ParsingUtils.sortList(vc.getFilters()))
: (filtersWereAppliedToContext || vc.filtersWereApplied()
? VCFConstants.PASSES_FILTERS_v4
: VCFConstants.UNFILTERED);
mWriter.write(filters);
mWriter.write(VCFConstants.FIELD_SEPARATOR);
// INFO
Map<String, String> infoFields = new TreeMap<String, String>();
for (Map.Entry<String, Object> field : vc.getAttributes().entrySet()) {
String key = field.getKey();
if (key.equals(VariantContext.ID_KEY)
|| key.equals(VariantContext.REFERENCE_BASE_FOR_INDEL_KEY)
|| key.equals(VariantContext.UNPARSED_GENOTYPE_MAP_KEY)
|| key.equals(VariantContext.UNPARSED_GENOTYPE_PARSER_KEY)) continue;
String outputValue = formatVCFField(field.getValue());
if (outputValue != null) infoFields.put(key, outputValue);
}
writeInfoString(infoFields);
// FORMAT
if (vc.hasAttribute(VariantContext.UNPARSED_GENOTYPE_MAP_KEY)) {
mWriter.write(VCFConstants.FIELD_SEPARATOR);
mWriter.write(vc.getAttributeAsString(VariantContext.UNPARSED_GENOTYPE_MAP_KEY, ""));
} else {
List<String> genotypeAttributeKeys = new ArrayList<String>();
if (vc.hasGenotypes()) {
genotypeAttributeKeys.addAll(calcVCFGenotypeKeys(vc));
} else if (mHeader.hasGenotypingData()) {
// this needs to be done in case all samples are no-calls
genotypeAttributeKeys.add(VCFConstants.GENOTYPE_KEY);
}
if (genotypeAttributeKeys.size() > 0) {
String genotypeFormatString =
ParsingUtils.join(VCFConstants.GENOTYPE_FIELD_SEPARATOR, genotypeAttributeKeys);
mWriter.write(VCFConstants.FIELD_SEPARATOR);
mWriter.write(genotypeFormatString);
addGenotypeData(vc, alleleMap, genotypeAttributeKeys);
}
}
mWriter.write("\n");
mWriter.flush(); // necessary so that writing to an output stream will work
} catch (IOException e) {
throw new RuntimeException("Unable to write the VCF object to " + locationString());
}
}
public static VariantContext createVariantContextWithTrimmedAlleles(VariantContext inputVC) {
// see if we need to trim common reference base from all alleles
boolean trimVC;
// We need to trim common reference base from all alleles in all genotypes if a ref base is
// common to all alleles
Allele refAllele = inputVC.getReference();
if (!inputVC.isVariant()) trimVC = false;
else if (refAllele.isNull()) trimVC = false;
else {
trimVC =
(AbstractVCFCodec.computeForwardClipping(
new ArrayList<Allele>(inputVC.getAlternateAlleles()),
inputVC.getReference().getDisplayString())
> 0);
}
// nothing to do if we don't need to trim bases
if (trimVC) {
List<Allele> alleles = new ArrayList<Allele>();
GenotypesContext genotypes = GenotypesContext.create();
// set the reference base for indels in the attributes
Map<String, Object> attributes = new TreeMap<String, Object>(inputVC.getAttributes());
Map<Allele, Allele> originalToTrimmedAlleleMap = new HashMap<Allele, Allele>();
for (Allele a : inputVC.getAlleles()) {
if (a.isSymbolic()) {
alleles.add(a);
originalToTrimmedAlleleMap.put(a, a);
} else {
// get bases for current allele and create a new one with trimmed bases
byte[] newBases = Arrays.copyOfRange(a.getBases(), 1, a.length());
Allele trimmedAllele = Allele.create(newBases, a.isReference());
alleles.add(trimmedAllele);
originalToTrimmedAlleleMap.put(a, trimmedAllele);
}
}
// detect case where we're trimming bases but resulting vc doesn't have any null allele. In
// that case, we keep original representation
// example: mixed records such as {TA*,TGA,TG}
boolean hasNullAlleles = false;
for (Allele a : originalToTrimmedAlleleMap.values()) {
if (a.isNull()) hasNullAlleles = true;
if (a.isReference()) refAllele = a;
}
if (!hasNullAlleles) return inputVC;
// now we can recreate new genotypes with trimmed alleles
for (final Genotype genotype : inputVC.getGenotypes()) {
List<Allele> originalAlleles = genotype.getAlleles();
List<Allele> trimmedAlleles = new ArrayList<Allele>();
for (Allele a : originalAlleles) {
if (a.isCalled()) trimmedAlleles.add(originalToTrimmedAlleleMap.get(a));
else trimmedAlleles.add(Allele.NO_CALL);
}
genotypes.add(Genotype.modifyAlleles(genotype, trimmedAlleles));
}
final VariantContextBuilder builder = new VariantContextBuilder(inputVC);
return builder
.alleles(alleles)
.genotypes(genotypes)
.attributes(attributes)
.referenceBaseForIndel(new Byte(inputVC.getReference().getBases()[0]))
.make();
}
return inputVC;
}
private List<Haplotype> computeHaplotypes(
final ReadBackedPileup pileup,
final int contextSize,
final int locus,
final VariantContext vc) {
// Compute all possible haplotypes consistent with current pileup
int haplotypesToCompute = vc.getAlternateAlleles().size() + 1;
final PriorityQueue<Haplotype> candidateHaplotypeQueue =
new PriorityQueue<Haplotype>(100, new HaplotypeComparator());
final PriorityQueue<Haplotype> consensusHaplotypeQueue =
new PriorityQueue<Haplotype>(
MAX_CONSENSUS_HAPLOTYPES_TO_CONSIDER, new HaplotypeComparator());
for (final PileupElement p : pileup) {
final Haplotype haplotypeFromRead = getHaplotypeFromRead(p, contextSize, locus);
candidateHaplotypeQueue.add(haplotypeFromRead);
}
// Now that priority queue has been built with all reads at context, we need to merge and find
// possible segregating haplotypes
Haplotype elem;
while ((elem = candidateHaplotypeQueue.poll()) != null) {
boolean foundHaplotypeMatch = false;
Haplotype lastCheckedHaplotype = null;
for (final Haplotype haplotypeFromList : consensusHaplotypeQueue) {
final Haplotype consensusHaplotype = getConsensusHaplotype(elem, haplotypeFromList);
if (consensusHaplotype != null) {
foundHaplotypeMatch = true;
if (consensusHaplotype.getQualitySum() > haplotypeFromList.getQualitySum()) {
consensusHaplotypeQueue.remove(haplotypeFromList);
consensusHaplotypeQueue.add(consensusHaplotype);
}
break;
} else {
lastCheckedHaplotype = haplotypeFromList;
}
}
if (!foundHaplotypeMatch
&& consensusHaplotypeQueue.size() < MAX_CONSENSUS_HAPLOTYPES_TO_CONSIDER) {
consensusHaplotypeQueue.add(elem);
} else if (!foundHaplotypeMatch
&& lastCheckedHaplotype != null
&& elem.getQualitySum() > lastCheckedHaplotype.getQualitySum()) {
consensusHaplotypeQueue.remove(lastCheckedHaplotype);
consensusHaplotypeQueue.add(elem);
}
}
// Now retrieve the N most popular haplotypes
if (consensusHaplotypeQueue.size() > 0) {
// The consensus haplotypes are in a quality-ordered priority queue, so the best haplotypes
// are just the ones at the front of the queue
final Haplotype haplotype1 = consensusHaplotypeQueue.poll();
List<Haplotype> hlist = new ArrayList<Haplotype>();
hlist.add(new Haplotype(haplotype1.getBases(), 60));
for (int k = 1; k < haplotypesToCompute; k++) {
Haplotype haplotype2 = consensusHaplotypeQueue.poll();
if (haplotype2 == null) {
haplotype2 = haplotype1;
} // Sometimes only the reference haplotype can be found
hlist.add(new Haplotype(haplotype2.getBases(), 20));
}
return hlist;
} else return null;
}
/**
* Main entry function to calculate genotypes of a given VC with corresponding GL's
*
* @param tracker Tracker
* @param refContext Reference context
* @param rawContext Raw context
* @param stratifiedContexts Stratified alignment contexts
* @param vc Input VC
* @param model GL calculation model
* @param inheritAttributesFromInputVC Output VC will contain attributes inherited from input vc
* @return VC with assigned genotypes
*/
public VariantCallContext calculateGenotypes(
final RefMetaDataTracker tracker,
final ReferenceContext refContext,
final AlignmentContext rawContext,
Map<String, AlignmentContext> stratifiedContexts,
final VariantContext vc,
final GenotypeLikelihoodsCalculationModel.Model model,
final boolean inheritAttributesFromInputVC,
final Map<String, org.broadinstitute.sting.utils.genotyper.PerReadAlleleLikelihoodMap>
perReadAlleleLikelihoodMap) {
boolean limitedContext =
tracker == null || refContext == null || rawContext == null || stratifiedContexts == null;
// initialize the data for this thread if that hasn't been done yet
if (afcm.get() == null) {
afcm.set(AFCalcFactory.createAFCalc(UAC, N, logger));
}
// estimate our confidence in a reference call and return
if (vc.getNSamples() == 0) {
if (limitedContext) return null;
return (UAC.OutputMode != OUTPUT_MODE.EMIT_ALL_SITES
? estimateReferenceConfidence(vc, stratifiedContexts, getTheta(model), false, 1.0)
: generateEmptyContext(tracker, refContext, stratifiedContexts, rawContext));
}
AFCalcResult AFresult = afcm.get().getLog10PNonRef(vc, getAlleleFrequencyPriors(model));
// is the most likely frequency conformation AC=0 for all alternate alleles?
boolean bestGuessIsRef = true;
// determine which alternate alleles have AF>0
final List<Allele> myAlleles = new ArrayList<Allele>(vc.getAlleles().size());
final List<Integer> alleleCountsofMLE = new ArrayList<Integer>(vc.getAlleles().size());
myAlleles.add(vc.getReference());
for (int i = 0; i < AFresult.getAllelesUsedInGenotyping().size(); i++) {
final Allele alternateAllele = AFresult.getAllelesUsedInGenotyping().get(i);
if (alternateAllele.isReference()) continue;
// we are non-ref if the probability of being non-ref > the emit confidence.
// the emit confidence is phred-scaled, say 30 => 10^-3.
// the posterior AF > 0 is log10: -5 => 10^-5
// we are non-ref if 10^-5 < 10^-3 => -5 < -3
final boolean isNonRef =
AFresult.isPolymorphic(alternateAllele, UAC.STANDARD_CONFIDENCE_FOR_EMITTING / -10.0);
// if the most likely AC is not 0, then this is a good alternate allele to use
if (isNonRef) {
myAlleles.add(alternateAllele);
alleleCountsofMLE.add(AFresult.getAlleleCountAtMLE(alternateAllele));
bestGuessIsRef = false;
}
// if in GENOTYPE_GIVEN_ALLELES mode, we still want to allow the use of a poor allele
else if (UAC.GenotypingMode
== GenotypeLikelihoodsCalculationModel.GENOTYPING_MODE.GENOTYPE_GIVEN_ALLELES) {
myAlleles.add(alternateAllele);
alleleCountsofMLE.add(AFresult.getAlleleCountAtMLE(alternateAllele));
}
}
final double PoFGT0 = Math.pow(10, AFresult.getLog10PosteriorOfAFGT0());
// note the math.abs is necessary because -10 * 0.0 => -0.0 which isn't nice
final double phredScaledConfidence =
Math.abs(
!bestGuessIsRef
|| UAC.GenotypingMode
== GenotypeLikelihoodsCalculationModel.GENOTYPING_MODE
.GENOTYPE_GIVEN_ALLELES
? -10 * AFresult.getLog10PosteriorOfAFEq0()
: -10 * AFresult.getLog10PosteriorOfAFGT0());
// return a null call if we don't pass the confidence cutoff or the most likely allele frequency
// is zero
if (UAC.OutputMode != OUTPUT_MODE.EMIT_ALL_SITES
&& !passesEmitThreshold(phredScaledConfidence, bestGuessIsRef)) {
// technically, at this point our confidence in a reference call isn't accurately estimated
// because it didn't take into account samples with no data, so let's get a better estimate
return limitedContext
? null
: estimateReferenceConfidence(vc, stratifiedContexts, getTheta(model), true, PoFGT0);
}
// start constructing the resulting VC
final GenomeLoc loc = genomeLocParser.createGenomeLoc(vc);
final VariantContextBuilder builder =
new VariantContextBuilder(
"UG_call", loc.getContig(), loc.getStart(), loc.getStop(), myAlleles);
builder.log10PError(phredScaledConfidence / -10.0);
if (!passesCallThreshold(phredScaledConfidence)) builder.filters(filter);
// create the genotypes
final GenotypesContext genotypes = afcm.get().subsetAlleles(vc, myAlleles, true, ploidy);
builder.genotypes(genotypes);
// print out stats if we have a writer
if (verboseWriter != null && !limitedContext)
printVerboseData(refContext.getLocus().toString(), vc, PoFGT0, phredScaledConfidence, model);
// *** note that calculating strand bias involves overwriting data structures, so we do that
// last
final HashMap<String, Object> attributes = new HashMap<String, Object>();
// inherit attributed from input vc if requested
if (inheritAttributesFromInputVC) attributes.putAll(vc.getAttributes());
// if the site was downsampled, record that fact
if (!limitedContext && rawContext.hasPileupBeenDownsampled())
attributes.put(VCFConstants.DOWNSAMPLED_KEY, true);
if (UAC.ANNOTATE_NUMBER_OF_ALLELES_DISCOVERED)
attributes.put(NUMBER_OF_DISCOVERED_ALLELES_KEY, vc.getAlternateAlleles().size());
// add the MLE AC and AF annotations
if (alleleCountsofMLE.size() > 0) {
attributes.put(VCFConstants.MLE_ALLELE_COUNT_KEY, alleleCountsofMLE);
final int AN = builder.make().getCalledChrCount();
final ArrayList<Double> MLEfrequencies = new ArrayList<Double>(alleleCountsofMLE.size());
// the MLEAC is allowed to be larger than the AN (e.g. in the case of all PLs being 0, the GT
// is ./. but the exact model may arbitrarily choose an AC>1)
for (int AC : alleleCountsofMLE) MLEfrequencies.add(Math.min(1.0, (double) AC / (double) AN));
attributes.put(VCFConstants.MLE_ALLELE_FREQUENCY_KEY, MLEfrequencies);
}
if (UAC.COMPUTE_SLOD && !limitedContext && !bestGuessIsRef) {
// final boolean DEBUG_SLOD = false;
// the overall lod
// double overallLog10PofNull = AFresult.log10AlleleFrequencyPosteriors[0];
double overallLog10PofF = AFresult.getLog10LikelihoodOfAFGT0();
// if ( DEBUG_SLOD ) System.out.println("overallLog10PofF=" + overallLog10PofF);
List<Allele> allAllelesToUse = builder.make().getAlleles();
// the forward lod
VariantContext vcForward =
calculateLikelihoods(
tracker,
refContext,
stratifiedContexts,
AlignmentContextUtils.ReadOrientation.FORWARD,
allAllelesToUse,
false,
model,
perReadAlleleLikelihoodMap);
AFresult = afcm.get().getLog10PNonRef(vcForward, getAlleleFrequencyPriors(model));
// double[] normalizedLog10Posteriors =
// MathUtils.normalizeFromLog10(AFresult.log10AlleleFrequencyPosteriors, true);
double forwardLog10PofNull = AFresult.getLog10LikelihoodOfAFEq0();
double forwardLog10PofF = AFresult.getLog10LikelihoodOfAFGT0();
// if ( DEBUG_SLOD ) System.out.println("forwardLog10PofNull=" + forwardLog10PofNull + ",
// forwardLog10PofF=" + forwardLog10PofF);
// the reverse lod
VariantContext vcReverse =
calculateLikelihoods(
tracker,
refContext,
stratifiedContexts,
AlignmentContextUtils.ReadOrientation.REVERSE,
allAllelesToUse,
false,
model,
perReadAlleleLikelihoodMap);
AFresult = afcm.get().getLog10PNonRef(vcReverse, getAlleleFrequencyPriors(model));
// normalizedLog10Posteriors =
// MathUtils.normalizeFromLog10(AFresult.log10AlleleFrequencyPosteriors, true);
double reverseLog10PofNull = AFresult.getLog10LikelihoodOfAFEq0();
double reverseLog10PofF = AFresult.getLog10LikelihoodOfAFGT0();
// if ( DEBUG_SLOD ) System.out.println("reverseLog10PofNull=" + reverseLog10PofNull + ",
// reverseLog10PofF=" + reverseLog10PofF);
double forwardLod = forwardLog10PofF + reverseLog10PofNull - overallLog10PofF;
double reverseLod = reverseLog10PofF + forwardLog10PofNull - overallLog10PofF;
// if ( DEBUG_SLOD ) System.out.println("forward lod=" + forwardLod + ", reverse lod=" +
// reverseLod);
// strand score is max bias between forward and reverse strands
double strandScore = Math.max(forwardLod, reverseLod);
// rescale by a factor of 10
strandScore *= 10.0;
// logger.debug(String.format("SLOD=%f", strandScore));
if (!Double.isNaN(strandScore)) attributes.put("SB", strandScore);
}
// finish constructing the resulting VC
builder.attributes(attributes);
VariantContext vcCall = builder.make();
// if we are subsetting alleles (either because there were too many or because some were not
// polymorphic)
// then we may need to trim the alleles (because the original VariantContext may have had to pad
// at the end).
if (myAlleles.size() != vc.getAlleles().size()
&& !limitedContext) // limitedContext callers need to handle allele trimming on their own to
// keep their perReadAlleleLikelihoodMap alleles in sync
vcCall = VariantContextUtils.reverseTrimAlleles(vcCall);
if (annotationEngine != null
&& !limitedContext) { // limitedContext callers need to handle annotations on their own by
// calling their own annotationEngine
// Note: we want to use the *unfiltered* and *unBAQed* context for the annotations
final ReadBackedPileup pileup = rawContext.getBasePileup();
stratifiedContexts = AlignmentContextUtils.splitContextBySampleName(pileup);
vcCall =
annotationEngine.annotateContext(
tracker, refContext, stratifiedContexts, vcCall, perReadAlleleLikelihoodMap);
}
return new VariantCallContext(vcCall, confidentlyCalled(phredScaledConfidence, PoFGT0));
}
private Map<String, Object> annotateIndel(AlignmentContext stratifiedContext, VariantContext vc) {
if (!stratifiedContext.hasExtendedEventPileup()) {
return null;
}
ReadBackedExtendedEventPileup pileup = stratifiedContext.getExtendedEventPileup();
if (pileup == null) return null;
int totalDepth = pileup.size();
Map<String, Object> map = new HashMap<String, Object>();
map.put(getKeyNames().get(0), totalDepth); // put total depth in right away
if (totalDepth == 0) return map;
int mq0 = 0; // number of "ref" reads that are acually mq0
HashMap<String, Integer> alleleCounts = new HashMap<String, Integer>();
Allele refAllele = vc.getReference();
for (Allele allele : vc.getAlternateAlleles()) {
if (allele.isNoCall()) {
continue; // this does not look so good, should we die???
}
alleleCounts.put(getAlleleRepresentation(allele), 0);
}
for (ExtendedEventPileupElement e : pileup.toExtendedIterable()) {
if (e.getMappingQual() == 0) {
mq0++;
continue;
}
if (e.isInsertion()) {
final String b = e.getEventBases();
if (alleleCounts.containsKey(b)) {
alleleCounts.put(b, alleleCounts.get(b) + 1);
}
} else {
if (e.isDeletion()) {
if (e.getEventLength() == refAllele.length()) {
// this is indeed the deletion allele recorded in VC
final String b = DEL;
if (alleleCounts.containsKey(b)) {
alleleCounts.put(b, alleleCounts.get(b) + 1);
}
}
// else {
// System.out.print(" deletion of WRONG length found");
// }
}
}
}
if (mq0 == totalDepth) return map;
String[] fracs = new String[alleleCounts.size()];
for (int i = 0; i < vc.getAlternateAlleles().size(); i++)
fracs[i] =
String.format(
"%.3f",
((float) alleleCounts.get(getAlleleRepresentation(vc.getAlternateAllele(i))))
/ (totalDepth - mq0));
map.put(getKeyNames().get(1), fracs);
// map.put(getKeyNames().get(0), counts);
return map;
}
