Revert AF calculation bug and yet another reblocking fix

src/main/java/org/broadinstitute/hellbender/tools/genomicsdb/GenomicsDBArgumentCollection.java

@@ -24,7 +24,7 @@ public class GenomicsDBArgumentCollection implements Serializable {
    * Must be at least one greater than the maximum number of alternate alleles for genotyping.
    * A typical value is 3 more than the --max-alternate-alleles value that's used by GenotypeGVCFs and larger differences
    * result in more robustness to PCR-related indel errors.
-   * Note that GenotypeGVCFs will drop highly multi-allelic sites that are missing likelihoods.
+   * NOTE: GenotypeGVCFs will drop multi-allelic sites with more than this many alternate alleles since they are missing likelihoods.
    *
    * See also {@link org.broadinstitute.hellbender.tools.walkers.genotyper.GenotypeCalculationArgumentCollection#MAX_ALTERNATE_ALLELES_LONG_NAME}
    */

src/main/java/org/broadinstitute/hellbender/tools/walkers/genotyper/AlleleSubsettingUtils.java

@@ -45,17 +45,13 @@ private AlleleSubsettingUtils() {}  // prevent instantiation
      * @param originalAlleles          the original alleles
      * @param allelesToKeep            the subset of alleles to use with the new Genotypes
      * @param assignmentMethod         assignment strategy for the (subsetted) PLs
-     * @param depth                    the original variant DP or 0 if there was no DP
-     * @param suppressUninformativePLs             force the output of a PL array even if there is no data
      * @return                         a new non-null GenotypesContext
      */
     public static GenotypesContext subsetAlleles(final GenotypesContext originalGs, final int defaultPloidy,
                                                  final List<Allele> originalAlleles,
                                                  final List<Allele> allelesToKeep,
                                                  final GenotypePriorCalculator gpc,
-                                                 final GenotypeAssignmentMethod assignmentMethod,
-                                                 final int depth,
-                                                 final boolean suppressUninformativePLs) {
+                                                 final GenotypeAssignmentMethod assignmentMethod) {
         Utils.nonNull(originalGs, "original GenotypesContext must not be null");
         Utils.nonNull(allelesToKeep, "allelesToKeep is null");
         Utils.nonEmpty(allelesToKeep, "must keep at least one allele");
@@ -94,9 +90,6 @@ public static GenotypesContext subsetAlleles(final GenotypesContext originalGs,
                 newLog10GQ = -0.1*g.getGQ();
             }
 
-            final boolean useNewLikelihoods = !suppressUninformativePLs ||
-                    (newLikelihoods != null && (depth != 0 || GATKVariantContextUtils.isInformative(newLikelihoods)));
-
             final GenotypeBuilder gb = new GenotypeBuilder(g);
             final Map<String, Object> attributes = new HashMap<>(g.getExtendedAttributes());
             attributes.remove(GATKVCFConstants.PHRED_SCALED_POSTERIORS_KEY);
@@ -107,9 +100,7 @@ public static GenotypesContext subsetAlleles(final GenotypesContext originalGs,
             if (newLog10GQ != Double.NEGATIVE_INFINITY && g.hasGQ()) {  //only put GQ if originally present
                 gb.log10PError(newLog10GQ);
             }
-            if (useNewLikelihoods) {
-                gb.PL(newLikelihoods);
-            }
+            gb.PL(newLikelihoods);
 
             GATKVariantContextUtils.makeGenotypeCall(g.getPloidy(), gb, assignmentMethod, newLikelihoods, allelesToKeep, g.getAlleles(), gpc);
 

src/main/java/org/broadinstitute/hellbender/tools/walkers/genotyper/GenotypingEngine.java

@@ -133,7 +133,8 @@ public VariantContext calculateGenotypes(final VariantContext vc, final Genotype
         if (maxAltAlleles < vc.getAlternateAlleles().size()) {
             final List<Allele> allelesToKeep = AlleleSubsettingUtils.calculateMostLikelyAlleles(vc, defaultPloidy, maxAltAlleles);
             final GenotypesContext reducedGenotypes = allelesToKeep.size() == 1 ? GATKVariantContextUtils.subsetToRefOnly(vc, defaultPloidy) :
-                    AlleleSubsettingUtils.subsetAlleles(vc.getGenotypes(), defaultPloidy, vc.getAlleles(), allelesToKeep, gpc, GenotypeAssignmentMethod.SET_TO_NO_CALL, vc.getAttributeAsInt(VCFConstants.DEPTH_KEY, 0), false);
+                    AlleleSubsettingUtils.subsetAlleles(vc.getGenotypes(), defaultPloidy, vc.getAlleles(), allelesToKeep, gpc,
+                            GenotypeAssignmentMethod.BEST_MATCH_TO_ORIGINAL);  //with no PLs in some reblocked GVCFs, no-calls are just going to cause problems, so keep 0/0 genotypes as such without trying to recall
             reducedVC = new VariantContextBuilder(vc).alleles(allelesToKeep).genotypes(reducedGenotypes).make();
         }
 
@@ -181,7 +182,7 @@ && noAllelesOrFirstAlleleIsNotNonRef(outputAlternativeAlleles.alleles) && givenA
         // create the genotypes
         //TODO: omit subsetting if output alleles is not a proper subset of vc.getAlleles
         final GenotypesContext genotypes = outputAlleles.size() == 1 ? GATKVariantContextUtils.subsetToRefOnly(vc, defaultPloidy) :
-                AlleleSubsettingUtils.subsetAlleles(vc.getGenotypes(), defaultPloidy, vc.getAlleles(), outputAlleles, gpc, configuration.genotypeArgs.genotypeAssignmentMethod, vc.getAttributeAsInt(VCFConstants.DEPTH_KEY, 0), false);
+                AlleleSubsettingUtils.subsetAlleles(vc.getGenotypes(), defaultPloidy, vc.getAlleles(), outputAlleles, gpc, configuration.genotypeArgs.genotypeAssignmentMethod);
 
         if (configuration.genotypeArgs.usePosteriorProbabilitiesToCalculateQual && hasPosteriors(genotypes)) {
             final double log10NoVariantPosterior = phredNoVariantPosteriorProbability(outputAlleles, genotypes) * -.1;
@@ -391,11 +392,11 @@ boolean isVcCoveredByDeletion(final VariantContext vc) {
      * {@link GenotypeLikelihoods#MAX_DIPLOID_ALT_ALLELES_THAT_CAN_BE_GENOTYPED}.
      */
     protected final boolean cannotBeGenotyped(final VariantContext vc) {
-        // protect against too many alternate alleles that we can't even run AF on:
         if (vc.getNAlleles() <= GenotypeLikelihoods.MAX_DIPLOID_ALT_ALLELES_THAT_CAN_BE_GENOTYPED
                 && vc.getGenotypes().stream().anyMatch(GenotypeUtils::genotypeIsUsableForAFCalculation)) {
             return false;
         }
+        // protect against too many alternate alleles that we can't even run AF on:
         if (vc.getNAlleles() > GenotypeLikelihoods.MAX_DIPLOID_ALT_ALLELES_THAT_CAN_BE_GENOTYPED) {
             logger.warn("Attempting to genotype more than " + GenotypeLikelihoods.MAX_DIPLOID_ALT_ALLELES_THAT_CAN_BE_GENOTYPED +
                     " alleles. Site will be skipped at location " + vc.getContig() + ":" + vc.getStart());

src/main/java/org/broadinstitute/hellbender/tools/walkers/genotyper/afcalc/AlleleFrequencyCalculator.java

@@ -77,7 +77,7 @@ private static double[] log10NormalizedGenotypePosteriors(final Genotype g, fina
         final double[] log10Likelihoods;
         if (g.hasLikelihoods()) {
             log10Likelihoods = g.getLikelihoods().getAsVector();
-        } else if (g.isHomRef()) {
+        } else if ( g.isHomRef()) {
             if (g.getPloidy() != 2) {
                 throw new IllegalStateException("Likelihoods are required to calculate posteriors for hom-refs with ploidy != 2, " +
                         "but were not found for genotype " + g + " with ploidy " + g.getPloidy());
@@ -88,6 +88,7 @@ private static double[] log10NormalizedGenotypePosteriors(final Genotype g, fina
                 throw new IllegalStateException("Genotype " + g + " does not contain GQ necessary to calculate posteriors.");
             }
         } else {
+            //no-call with no PLs are too risky -- don't assume they're reblocked hom-refs
             throw new IllegalStateException("Genotype " + g + " does not contain likelihoods necessary to calculate posteriors.");
         }
         final double[] log10Posteriors = new IndexRange(0, glCalc.genotypeCount()).mapToDouble(genotypeIndex -> {

src/main/java/org/broadinstitute/hellbender/tools/walkers/gnarlyGenotyper/GnarlyGenotyper.java

@@ -174,7 +174,7 @@ protected List<SimpleInterval> transformTraversalIntervals(final List<SimpleInte
     protected GenomicsDBOptions getGenomicsDBOptions() {
         if (genomicsDBOptions == null) {
             genomicsdbArgs.callGenotypes = CALL_GENOTYPES;
-            genotypeArgs.maxAlternateAlleles = PIPELINE_MAX_ALT_COUNT;
+            genomicsdbArgs.maxDiploidAltAllelesThatCanBeGenotyped = genotypeArgs.maxAlternateAlleles + 1; //plus one for internal NON_REF
             genomicsDBOptions = new GenomicsDBOptions(referenceArguments.getReferencePath(), genomicsdbArgs, genotypeArgs);
         }
         return genomicsDBOptions;

src/main/java/org/broadinstitute/hellbender/tools/walkers/variantutils/ReblockGVCF.java

@@ -512,9 +512,7 @@ protected GenotypeBuilder changeCallToHomRefVersusNonRef(final VariantContext lo
                 // the called alleles and this is a reference genotype that will stay hom-ref
                 final GenotypesContext context = AlleleSubsettingUtils.subsetAlleles(lowQualVariant.getGenotypes(),
                         genotype.getPloidy(), lowQualVariant.getAlleles(), Arrays.asList(inputRefAllele, bestAlt),
-                        null, GenotypeAssignmentMethod.BEST_MATCH_TO_ORIGINAL, //BEST_MATCH to avoid no-calling low qual genotypes
-                        lowQualVariant.getAttributeAsInt(VCFConstants.DEPTH_KEY, 0),
-                        false);  //emitEmptyPLs = true to make sure we always subset
+                        null, GenotypeAssignmentMethod.BEST_MATCH_TO_ORIGINAL);  //BEST_MATCH to avoid no-calling low qual genotypes
                 final Genotype subsetG = context.get(0);
                 gb = new GenotypeBuilder(subsetG).noAttributes();  //remove attributes because hom ref blocks shouldn't have posteriors
                 //subsetting may strip GQ and PLs for low qual genotypes
@@ -568,8 +566,7 @@ VariantContext cleanUpHighQualityVariant(final VariantContext variant) {
         if(allelesNeedSubsetting && !keepAllAlts) {
             newAlleleSetUntrimmed.removeAll(allelesToDrop);
             final GenotypesContext gc = AlleleSubsettingUtils.subsetAlleles(variant.getGenotypes(), genotype.getPloidy(), variant.getAlleles(),
-                    newAlleleSetUntrimmed, null, GenotypeAssignmentMethod.USE_PLS_TO_ASSIGN,
-                    variant.getAttributeAsInt(VCFConstants.DEPTH_KEY, 0), true);
+                    newAlleleSetUntrimmed, null, GenotypeAssignmentMethod.USE_PLS_TO_ASSIGN);
             if (gc.get(0).isHomRef() || !gc.get(0).hasGQ() || gc.get(0).getAlleles().contains(Allele.NO_CALL)) {  //could be low quality or no-call after subsetting
                 if (dropLowQuals) {
                     return null;
@@ -780,8 +777,8 @@ private static void addQualAnnotations(final Map<String, Object> destination, fi
                 //TODO: this isn't going to work for DRAGEN's genotype posteriors
                 final GenotypesContext gc = AlleleSubsettingUtils.subsetAlleles(updatedAllelesVC.getGenotypes(),
                         updatedAllelesGenotype.getPloidy(), updatedAllelesVC.getAlleles(), Arrays.asList(updatedAllelesVC.getReference(), alt), null,
-                        GenotypeAssignmentMethod.BEST_MATCH_TO_ORIGINAL, 0, true);
-                //assignment method doens't really matter as long as we don't zero out PLs; don't need depth to get PLs for quals
+                        GenotypeAssignmentMethod.BEST_MATCH_TO_ORIGINAL);
+                //assignment method doesn't really matter as long as we don't zero out PLs; don't need depth to get PLs for quals
 
                 final Genotype subsettedGenotype = gc.get(0);
                 final int[] likelihoods = getGenotypePosteriorsOtherwiseLikelihoods(subsettedGenotype, posteriorsKey);
@@ -862,8 +859,10 @@ private static void copyInfoAnnotations(final Map<String, Object> destinationAtt
                             final List<String> subsetList;
                             if (alleleSpecificValues.size() > 0) {
                                 subsetList = AlleleSubsettingUtils.remapRLengthList(alleleSpecificValues, relevantIndices, "");
-                                //zero out non-ref value, just in case
-                                subsetList.set(subsetList.size()-1,((AlleleSpecificAnnotation)annotation).getEmptyRawValue());
+                                if (sourceVC.getAlleles().get(relevantIndices[relevantIndices.length - 1]).equals(Allele.NON_REF_ALLELE)) {
+                                    //zero out non-ref value, just in case
+                                    subsetList.set(subsetList.size() - 1, ((AlleleSpecificAnnotation) annotation).getEmptyRawValue());
+                                }
                             } else {
                                 subsetList = Collections.nCopies(relevantIndices.length, "");
                             }

src/main/java/org/broadinstitute/hellbender/tools/walkers/variantutils/SelectVariants.java

@@ -1032,8 +1032,7 @@ private VariantContext subsetRecord(final VariantContext vc, final boolean prese
             final GenotypesContext subGenotypesWithOldAlleles = sub.getGenotypes();  //we need sub for the right samples, but PLs still go with old alleles
             newGC = sub.getNAlleles() == vc.getNAlleles() ? subGenotypesWithOldAlleles :
                     AlleleSubsettingUtils.subsetAlleles(subGenotypesWithOldAlleles, 0, vc.getAlleles(),
-                            sub.getAlleles(), null, GenotypeAssignmentMethod.DO_NOT_ASSIGN_GENOTYPES,
-                            vc.getAttributeAsInt(VCFConstants.DEPTH_KEY, 0), false);
+                            sub.getAlleles(), null, GenotypeAssignmentMethod.DO_NOT_ASSIGN_GENOTYPES);
         } else {
             newGC = sub.getGenotypes();
         }

src/main/java/org/broadinstitute/hellbender/utils/GenotypeUtils.java

@@ -139,6 +139,7 @@ public static GenotypeCounts computeDiploidGenotypeCounts(final VariantContext v
     /**
      * Do we have (or can we infer) likelihoods necessary for allele frequency calculation?
      * Some reblocked and/or DRAGEN GVCFs omit likelihoods for ref blocks, but we can estimate them
+     * If GenomicsDB max alt threshold is too low, non-reference genotypes may also be missing PLs -- we can't estimate, so reject those
      * @param g a genotype of unknown call and ploidy
      * @return  true if we have enough info for AF calculation
      */

src/main/java/org/broadinstitute/hellbender/utils/variant/GATKVariantContextUtils.java

@@ -1687,7 +1687,7 @@ public static List<VariantContext> splitVariantContextToBiallelics(final Variant
                         genotypeAssignmentMethodUsed != GenotypeAssignmentMethod.SET_TO_NO_CALL)
                     AlleleSubsettingUtils.addInfoFieldAnnotations(vc, builder, keepOriginalChrCounts);
 
-                builder.genotypes(AlleleSubsettingUtils.subsetAlleles(vc.getGenotypes(),2,vc.getAlleles(), alleles, null, genotypeAssignmentMethodUsed,vc.getAttributeAsInt("DP",0), true));
+                builder.genotypes(AlleleSubsettingUtils.subsetAlleles(vc.getGenotypes(),2,vc.getAlleles(), alleles, null, genotypeAssignmentMethodUsed));
                 final VariantContext trimmed = trimAlleles(builder.make(), trimLeft, true);
                 biallelics.add(trimmed);
             }

src/test/java/org/broadinstitute/hellbender/tools/walkers/GenotypeGVCFsIntegrationTest.java

@@ -156,7 +156,12 @@ public Object[][] gvcfsToGenotype() {
                 {getTestFile( "combined.single.sample.pipeline.gatk3.vcf"),
                         getTestFile( "expected/includeLowQualSites.vcf"),
                         Arrays.asList( " --" + GenotypeGVCFs.ALL_SITES_LONG_NAME + " -L 20:10,012,730-10,012,740"),
-                        b37_reference_20_21}
+                        b37_reference_20_21},
+
+                //23 highly multi-allelic sites across 54 1000G exomes to test allele subsetting and QUAL calculation
+                {getTestFile("multiallelicQualRegression.vcf "),
+                        getTestFile("multiallelicQualRegression.expected.vcf"),
+                        NO_EXTRA_ARGS, hg38Reference}
         };
     }
 
@@ -281,23 +286,35 @@ public void assertMatchingGenotypesFromGenomicsDB(File input, File expected, Loc
         runGenotypeGVCFSAndAssertComparison(genomicsDBUri, expected, args, VariantContextTestUtils::assertVariantContextsHaveSameGenotypes, reference);
     }
 
-    @Test
-    public void testMaxAltsToCombineInGenomicsDB() {
-        final File tempGenomicsDB = GenomicsDBTestUtils.createTempGenomicsDB(CEUTRIO_20_21_GATK3_4_G_VCF, new SimpleInterval("20", 1, 11_000_000));
-        final String genomicsDBUri = GenomicsDBTestUtils.makeGenomicsDBUri(tempGenomicsDB);
-        final List<String> args = new ArrayList<String>();
+    @Test  //here GDBMaxAlts is greater than GGVCFsMaxAlts
+    public void testMaxAltsToCombineInGenomicsDB() throws IOException {
+        //multi-input tests
+        //8 ALT VC will get dropped if GDB max is < 8 because GDB doesn't return PLs and GGVCFs drops variants with no PLs
+        final String gnarlyTestPath = toolsTestDir + "walkers/GnarlyGenotyper/";
+        final List<File> inputs = Arrays.asList(new File(gnarlyTestPath + "sample6.vcf"),
+                new File(gnarlyTestPath + "sample7.vcf"),
+                new File(gnarlyTestPath + "sample8.vcf"),
+                new File(gnarlyTestPath + "sample9.vcf"));
+        final SimpleInterval interval =  new SimpleInterval("chr20", 257008, 257008);
+        final File tempGenomicsDB2 = GenomicsDBTestUtils.createTempGenomicsDB(inputs, interval);
+        final String genomicsDBUri2 = GenomicsDBTestUtils.makeGenomicsDBUri(tempGenomicsDB2);
+        final List<String> args = new ArrayList<>();
+        args.add("--"+GenomicsDBArgumentCollection.MAX_ALTS_LONG_NAME);
+        args.add("7");
         args.add("--"+GenotypeCalculationArgumentCollection.MAX_ALTERNATE_ALLELES_LONG_NAME);
-        args.add("3");
-        args.add("--" + GenomicsDBArgumentCollection.MAX_ALTS_LONG_NAME);
-        args.add("4");
-        runGenotypeGVCFSAndAssertCount(genomicsDBUri, args, 3, VariantContextTestUtils::assertVariantContextMaxAltAlleleCount, b37_reference_20_21);
+        args.add("5");
+        final File output = runGenotypeGVCFS(genomicsDBUri2, null, args, hg38Reference);
+        final Pair<VCFHeader, List<VariantContext>> outputDataNoVariant = VariantContextTestUtils.readEntireVCFIntoMemory(output.getAbsolutePath());
+        Assert.assertTrue(outputDataNoVariant.getRight().isEmpty());
 
-        args.clear();
+        //8 ALT VC will be output if GDB max is >= 8, but with only as many ALTs are requested in the GenotypeCalculationArguments
+        final List<String> args2 = new ArrayList<String>();
+        args.add("--"+GenomicsDBArgumentCollection.MAX_ALTS_LONG_NAME);
+        args.add("15");
         args.add("--"+GenotypeCalculationArgumentCollection.MAX_ALTERNATE_ALLELES_LONG_NAME);
-        args.add("2");
-        args.add("--" + GenomicsDBArgumentCollection.MAX_ALTS_LONG_NAME);
-        args.add("20");
-        runGenotypeGVCFSAndAssertCount(genomicsDBUri, args, 2, VariantContextTestUtils::assertVariantContextMaxAltAlleleCount, b37_reference_20_21);
+        args.add("5");
+        runGenotypeGVCFSAndAssertComparison(genomicsDBUri2, getTestFile("fourSamplesEightAlts.expected.vcf"), args2,
+                VariantContextTestUtils::assertVariantContextsHaveSameGenotypes, hg38Reference);
     }
 
     @Test(expectedExceptions = UserException.BadInput.class)
@@ -333,7 +350,7 @@ private void runAndCheckGenomicsDBOutput(final ArgumentsBuilder args, final File
         runCommandLine(args);
 
         // Note that if this isn't working it will take *FOREVER*
-        // runs in 0.06 minutes with no input intervals specfied
+        // runs in 0.06 minutes with no input intervals specified
         final List<VariantContext> expectedVC = VariantContextTestUtils.getVariantContexts(expected);
         final List<VariantContext> actualVC = VariantContextTestUtils.getVariantContexts(output);
         assertForEachElementInLists(actualVC, expectedVC, VariantContextTestUtils::assertVariantContextsHaveSameGenotypes);
@@ -422,6 +439,10 @@ private void runGenotypeGVCFSAndAssertComparison(File input, File expected, List
         );
     }
 
+
+    /**
+     * Note that this method does not use expected for comparison, but rather for updating exact match outputs
+     */
     private File runGenotypeGVCFS(String input, File expected, List<String> additionalArguments, String reference) {
         final File output = UPDATE_EXACT_MATCH_EXPECTED_OUTPUTS ? expected : createTempFile("genotypegvcf", ".vcf");
         final ArgumentsBuilder args = new ArgumentsBuilder();