Examples with VDJCAlignments com.milaboratory.mixcr.basictypes.VDJCAlignments used on opensource projects

Example 16 with VDJCAlignments

use of com.milaboratory.mixcr.basictypes.VDJCAlignments in project mixcr by milaboratory.

the class FieldExtractorsTest method testAnchorPoints1.

@Test
public void testAnchorPoints1() throws Exception {
    final boolean print = false;
    final Well44497b rg = new Well44497b(12312);
    final VDJCAlignerParameters rnaSeqParams = VDJCParametersPresets.getByName("rna-seq");
    final PartialAlignmentsAssemblerAligner aligner = new PartialAlignmentsAssemblerAligner(rnaSeqParams);
    final VDJCLibrary lib = VDJCLibraryRegistry.getDefault().getLibrary("default", "hs");
    for (VDJCGene gene : VDJCLibraryRegistry.getDefault().getLibrary("default", "hs").getGenes()) if (gene.isFunctional())
        aligner.addGene(gene);
    final TargetBuilder.VDJCGenes genes = new TargetBuilder.VDJCGenes(lib, "TRBV12-3*00", "TRBD1*00", "TRBJ1-3*00", "TRBC2*00");
    // | 310  | 338   | 438
    // 250V + 60CDR3 (20V 7N 10D 3N 20J) + 28J + 100C + 100N
    // "{CDR3Begin(-250)}V*270 NNNNNNN {DBegin(0)}D*10 NNN {CDR3End(-20):FR4End} {CBegin}C*100 N*100"
    final FieldExtractors.ExtractDefaultReferencePointsPositions extractor = new FieldExtractors.ExtractDefaultReferencePointsPositions();
    F6 goAssert = new F6() {

        @Override
        public Integer[][] go(String seq, int len, int offset1, int offset2, int offset3, String expected) {
            final NucleotideSequence baseSeq = TargetBuilder.generateSequence(genes, seq, rg);
            NucleotideSequence seq1 = baseSeq.getRange(offset1, Math.min(baseSeq.size(), offset1 + len));
            NucleotideSequence seq2 = offset2 == -1 ? null : baseSeq.getRange(offset2, Math.min(baseSeq.size(), offset2 + len));
            NucleotideSequence seq3 = offset3 == -1 ? null : baseSeq.getRange(offset3, Math.min(baseSeq.size(), offset3 + len));
            VDJCAlignmentResult<VDJCMultiRead> alignment = offset3 == -1 ? offset2 == -1 ? aligner.process(MiXCRTestUtils.createMultiRead(seq1)) : aligner.process(MiXCRTestUtils.createMultiRead(seq1, seq2)) : aligner.process(MiXCRTestUtils.createMultiRead(seq1, seq2, seq3));
            VDJCAlignments al = alignment.alignment;
            Assert.assertNotNull(al);
            if (print) {
                MiXCRTestUtils.printAlignment(al);
                System.out.println();
                System.out.println("-------------------------------------------");
                System.out.println();
            }
            String val = extractor.extract(al);
            if (print)
                System.out.println(val);
            String[] spl = val.split(",");
            Integer[][] result = new Integer[spl.length][ReferencePoint.DefaultReferencePoints.length];
            for (int i = 0; i < spl.length; i++) {
                String[] spl1 = spl[i].split(":");
                for (int j = 0; j < spl1.length; j++) {
                    try {
                        result[i][j] = Integer.decode(spl1[j]);
                    } catch (NumberFormatException e) {
                    }
                }
            }
            return result;
        }
    };
    // No PSegments, just deletions
    Integer[][] r = goAssert.go("{CDR3Begin(-250):VEnd(-3)} 'CCAAA' {DBegin(0):DEnd(0)} 'AAA' {JBegin(2):FR4End} " + "{CBegin}C*100 N*100", 100, 240, 307, 450, "");
    assertExportPoint(r[0], ReferencePoint.VEnd, -3);
    assertExportPoint(r[0], ReferencePoint.DBegin, 0);
    assertExportPoint(r[0], ReferencePoint.DEnd, 0);
    assertExportPoint(r[0], ReferencePoint.JBegin, -2);
    r = goAssert.go("{CDR3Begin(-250):VEnd(0)} 'CCAAA' {DBegin(0):DEnd(-2)} 'AAA' {JBegin:FR4End} {CBegin}C*100 N*100", 100, 240, 307, 450, "");
    assertExportPoint(r[0], ReferencePoint.VEnd, 0);
    assertExportPoint(r[0], ReferencePoint.DBegin, 0);
    assertExportPoint(r[0], ReferencePoint.DEnd, -2);
    assertExportPoint(r[0], ReferencePoint.JBegin, 0);
    // With PSegments
    r = goAssert.go("{CDR3Begin(-250):VEnd(0)} {VEnd:VEnd(-3)} 'CCAAA' {DBegin(3):DBegin} {DBegin:DEnd(-2)} 'AAA' " + "{JBegin(2):JBegin} {JBegin:FR4End} {CBegin}C*100 N*100", 100, 240, 307, 450, "");
    assertExportPoint(r[0], ReferencePoint.VEnd, 3);
    assertExportPoint(r[0], ReferencePoint.DBegin, 3);
    assertExportPoint(r[0], ReferencePoint.DEnd, -2);
    assertExportPoint(r[0], ReferencePoint.JBegin, 2);
}

Example 17 with VDJCAlignments

use of com.milaboratory.mixcr.basictypes.VDJCAlignments in project mixcr by milaboratory.

the class FullSeqAssemblerTest method testRandom1.

@Test
public void testRandom1() throws Exception {
    CloneFraction[] clones = { new CloneFraction(750, masterSeq1WT), // V: S346:G->T
    new CloneFraction(1000, masterSeq1VSub1), // J: D55:A
    new CloneFraction(1000, masterSeq1VDel1JDel1), // J: D62:C
    new CloneFraction(500, masterSeq1VDel1JDelVSub2) };
    Well19937c rand = new Well19937c();
    rand.setSeed(12345);
    RandomDataGenerator rdg = new RandomDataGenerator(rand);
    List<SequenceRead> readsOrig = new ArrayList<>();
    int readLength = 100;
    int id = -1;
    for (CloneFraction clone : clones) {
        for (int i = 0; i < clone.count; i++) {
            // Left read with CDR3
            ++id;
            readsOrig.add(new PairedRead(new SingleReadImpl(id, new NSequenceWithQuality(clone.seq.getRangeFromCDR3Begin(-rand.nextInt(readLength - clone.seq.cdr3Part), readLength)), "R1_" + id), new SingleReadImpl(id, new NSequenceWithQuality(clone.seq.getRangeFromCDR3End(rdg.nextInt(-clone.seq.cdr3Part / 2, clone.seq.jPart), readLength).getReverseComplement()), "R2_" + id)));
            ++id;
            readsOrig.add(new PairedRead(new SingleReadImpl(id, new NSequenceWithQuality(clone.seq.getRangeFromCDR3Begin(rdg.nextInt(-clone.seq.vPart, clone.seq.cdr3Part / 2 - readLength), readLength)), "R1_" + id), new SingleReadImpl(id, new NSequenceWithQuality(clone.seq.getRangeFromCDR3Begin(-rand.nextInt(readLength - clone.seq.cdr3Part), readLength)).getReverseComplement(), "R2_" + id)));
        }
    }
    // readsOrig = Arrays.asList(setReadId(0, readsOrig.get(12)), setReadId(1, readsOrig.get(13)));
    int[] perm = rdg.nextPermutation(readsOrig.size(), readsOrig.size());
    List<SequenceRead> reads = new ArrayList<>();
    for (int i = 0; i < readsOrig.size(); i++) reads.add(readsOrig.get(perm[i]));
    RunMiXCR.RunMiXCRAnalysis params = new RunMiXCR.RunMiXCRAnalysis(new SequenceReaderCloseable<SequenceRead>() {

        int counter = 0;

        @Override
        public void close() {
        }

        @Override
        public long getNumberOfReads() {
            return counter;
        }

        @Override
        public synchronized SequenceRead take() {
            if (counter == reads.size())
                return null;
            return reads.get(counter++);
        }
    }, true);
    params.alignerParameters = VDJCParametersPresets.getByName("rna-seq");
    params.alignerParameters.setSaveOriginalReads(true);
    params.alignerParameters.setVAlignmentParameters(params.alignerParameters.getVAlignerParameters().setGeneFeatureToAlign(GeneFeature.VTranscriptWithP));
    RunMiXCR.AlignResult align = RunMiXCR.align(params);
    // // TODO exception for translation
    // for (VDJCAlignments al : align.alignments) {
    // for (int i = 0; i < al.numberOfTargets(); i++) {
    // System.out.println(VDJCAlignmentsFormatter.getTargetAsMultiAlignment(al, i));
    // System.out.println();
    // }
    // System.out.println();
    // System.out.println(" ================================================ ");
    // System.out.println();
    // }
    RunMiXCR.AssembleResult assemble = RunMiXCR.assemble(align);
    Assert.assertEquals(1, assemble.cloneSet.size());
    CloneFactory cloneFactory = new CloneFactory(align.parameters.cloneAssemblerParameters.getCloneFactoryParameters(), align.parameters.cloneAssemblerParameters.getAssemblingFeatures(), align.usedGenes, align.parameters.alignerParameters.getFeaturesToAlignMap());
    FullSeqAssembler agg = new FullSeqAssembler(cloneFactory, DEFAULT_PARAMETERS, assemble.cloneSet.get(0), align.parameters.alignerParameters);
    FullSeqAssembler.RawVariantsData prep = agg.calculateRawData(() -> CUtils.asOutputPort(align.alignments.stream().filter(a -> a.getFeature(GeneFeature.CDR3) != null).collect(Collectors.toList())));
    List<Clone> clns = new ArrayList<>(new CloneSet(Arrays.asList(agg.callVariants(prep))).getClones());
    clns.sort(Comparator.comparingDouble(Clone::getCount).reversed());
    System.out.println("# Clones: " + clns.size());
    id = 0;
    for (Clone clone : clns) {
        clone = clone.setId(id++);
        System.out.println(clone.numberOfTargets());
        System.out.println(clone.getCount());
        System.out.println(clone.getFraction());
        System.out.println(clone.getBestHit(GeneType.Variable).getAlignment(0).getAbsoluteMutations());
        System.out.println(clone.getBestHit(GeneType.Joining).getAlignment(0).getAbsoluteMutations());
        System.out.println();
    // ActionExportClonesPretty.outputCompact(System.out, clone);
    }
}

Example 18 with VDJCAlignments

use of com.milaboratory.mixcr.basictypes.VDJCAlignments in project mixcr by milaboratory.

the class FullSeqAssemblerTest method test1.

@Test
public void test1() throws Exception {
    int len = 140;
    PairedRead read1 = new PairedRead(new SingleReadImpl(0, new NSequenceWithQuality(masterSeq1WT.getRangeFromCDR3Begin(-20, len)), "R1"), new SingleReadImpl(0, new NSequenceWithQuality(masterSeq1WT.getRangeFromCDR3Begin(-200, len).getReverseComplement()), "R2"));
    PairedRead read2 = new PairedRead(new SingleReadImpl(1, new NSequenceWithQuality(masterSeq1WT.getRangeFromCDR3Begin(-30, len)), "R1"), new SingleReadImpl(1, new NSequenceWithQuality(masterSeq1WT.getRangeFromCDR3Begin(-150, len).getReverseComplement()), "R2"));
    RunMiXCR.RunMiXCRAnalysis params = new RunMiXCR.RunMiXCRAnalysis(read1, read2);
    // [-200, -60]  [-20, 120]
    // [-150, 110]
    // 
    // [-200, -150], [110, 120] = 60
    // [-60, -20] = 40
    params.alignerParameters = VDJCParametersPresets.getByName("rna-seq");
    params.alignerParameters.setSaveOriginalReads(true);
    params.cloneAssemblerParameters.updateFrom(params.alignerParameters);
    RunMiXCR.AlignResult align = RunMiXCR.align(params);
    // for (VDJCAlignments al : align.alignments) {
    // for (int i = 0; i < al.numberOfTargets(); i++)
    // System.out.println(VDJCAlignmentsFormatter.getTargetAsMultiAlignment(al, i));
    // System.out.println();
    // }
    RunMiXCR.AssembleResult assemble = RunMiXCR.assemble(align);
    CloneFactory cloneFactory = new CloneFactory(align.parameters.cloneAssemblerParameters.getCloneFactoryParameters(), align.parameters.cloneAssemblerParameters.getAssemblingFeatures(), align.usedGenes, align.parameters.alignerParameters.getFeaturesToAlignMap());
    FullSeqAssembler agg = new FullSeqAssembler(cloneFactory, DEFAULT_PARAMETERS, assemble.cloneSet.get(0), align.parameters.alignerParameters);
    PointSequence[] r2s = agg.toPointSequences(align.alignments.get(1));
    TIntHashSet p2 = new TIntHashSet(Arrays.stream(r2s).mapToInt(s -> s.point).toArray());
    Assert.assertEquals(261 - masterSeq1WT.cdr3Part, p2.size());
    PointSequence[] r1s = agg.toPointSequences(align.alignments.get(0));
    TIntHashSet p1 = new TIntHashSet(Arrays.stream(r1s).mapToInt(s -> s.point).toArray());
    Assert.assertEquals(281 - masterSeq1WT.cdr3Part, p1.size());
    FullSeqAssembler.RawVariantsData prep = agg.calculateRawData(() -> CUtils.asOutputPort(align.alignments));
    long uniq1 = StreamSupport.stream(CUtils.it(prep.createPort()).spliterator(), false).mapToInt(l -> l[0]).filter(c -> c == 0xFFFFFFFF).count();
    long uniq2 = StreamSupport.stream(CUtils.it(prep.createPort()).spliterator(), false).mapToInt(l -> l[1]).filter(c -> c == 0xFFFFFFFF).count();
    Assert.assertEquals(40, uniq1);
    Assert.assertEquals(60, uniq2);
    for (Clone clone : new CloneSet(Arrays.asList(agg.callVariants(prep))).getClones()) {
        ActionExportClonesPretty.outputCompact(System.out, clone);
        System.out.println();
        System.out.println(" ================================================ ");
        System.out.println();
    }
}

Example 19 with VDJCAlignments

use of com.milaboratory.mixcr.basictypes.VDJCAlignments in project mixcr by milaboratory.

the class FullSeqAssemblerTest method testLargeCloneNoMismatches.

@Test
public void testLargeCloneNoMismatches() throws Exception {
    MasterSequence master = FullSeqAssemblerTest.masterSeq1WT;
    NSequenceWithQuality seq = new NSequenceWithQuality(master.getRange(-master.vPart + 10, 80), SequenceQuality.GOOD_QUALITY_VALUE);
    RunMiXCR.RunMiXCRAnalysis params0 = new RunMiXCR.RunMiXCRAnalysis(new SingleReadImpl(0, seq, ""));
    params0.cloneAssemblerParameters.setAssemblingFeatures(new GeneFeature[] { GeneFeature.VDJRegion });
    Clone largeClone = RunMiXCR.assemble(RunMiXCR.align(params0)).cloneSet.get(0);
    // ActionExportClonesPretty.outputCompact(System.out, largeClone);
    // System.exit(0);
    Well44497b rnd = new Well44497b(1234567889L);
    int nReads = 100_000;
    int readLength = 75, readGap = 150;
    // slice seq randomly
    PairedRead[] slicedReads = new PairedRead[nReads];
    for (int i = 0; i < nReads; ++i) {
        int r1from = rnd.nextInt(seq.size() - readLength - 1), r1to = r1from + readLength, r2from = r1from + 1 + rnd.nextInt(seq.size() - r1from - readLength - 1), r2to = r2from + readLength;
        assert r2from > r1from;
        slicedReads[i] = new PairedRead(new SingleReadImpl(i, seq.getRange(r1from, r1to), "" + i), new SingleReadImpl(i, seq.getRange(r2from, r2to).getReverseComplement(), "" + i));
    }
    RunMiXCR.RunMiXCRAnalysis params = new RunMiXCR.RunMiXCRAnalysis(slicedReads);
    // params.alignerParameters = VDJCParametersPresets.getByName("rna-seq");
    params.alignerParameters.setSaveOriginalReads(true);
    RunMiXCR.AlignResult align = RunMiXCR.align(params);
    RunMiXCR.AssembleResult assemble = RunMiXCR.assemble(align);
    // for (VDJCAlignments al : align.alignments) {
    // for (int i = 0; i < al.numberOfTargets(); i++) {
    // System.out.println(VDJCAlignmentsFormatter.getTargetAsMultiAlignment(al, i));
    // System.out.println();
    // }
    // System.out.println();
    // System.out.println(" ================================================ ");
    // System.out.println();
    // }
    Assert.assertEquals(1, assemble.cloneSet.size());
    Clone initialClone = assemble.cloneSet.get(0);
    NSequenceWithQuality cdr3 = initialClone.getFeature(GeneFeature.CDR3);
    List<VDJCAlignments> alignments = align.alignments.stream().filter(al -> cdr3.equals(al.getFeature(GeneFeature.CDR3))).collect(Collectors.toList());
    alignments.stream().filter(al -> Arrays.stream(al.getBestHit(GeneType.Variable).getAlignments()).filter(Objects::nonNull).anyMatch(a -> !a.getAbsoluteMutations().isEmpty())).filter(al -> al.getBestHit(GeneType.Variable).getGene().getName().contains("3-74")).forEach(al -> {
        for (int i = 0; i < al.numberOfTargets(); i++) {
            System.out.println(VDJCAlignmentsFormatter.getTargetAsMultiAlignment(al, i));
            System.out.println();
        }
        System.out.println();
        System.out.println(" ================================================ ");
        System.out.println();
    });
    // System.exit(0);
    System.out.println("=> Agg");
    CloneFactory cloneFactory = new CloneFactory(align.parameters.cloneAssemblerParameters.getCloneFactoryParameters(), align.parameters.cloneAssemblerParameters.getAssemblingFeatures(), align.usedGenes, align.parameters.alignerParameters.getFeaturesToAlignMap());
    FullSeqAssembler agg = new FullSeqAssembler(cloneFactory, DEFAULT_PARAMETERS, initialClone, align.parameters.alignerParameters);
    FullSeqAssembler.RawVariantsData prep = agg.calculateRawData(() -> CUtils.asOutputPort(alignments));
    List<Clone> clones = new ArrayList<>(new CloneSet(Arrays.asList(agg.callVariants(prep))).getClones());
    clones.sort(Comparator.comparingDouble(Clone::getCount).reversed());
    for (Clone clone : clones) {
        ActionExportClonesPretty.outputCompact(System.out, clone);
        System.out.println();
        System.out.println(" ================================================ ");
        System.out.println();
    }
}

Example 20 with VDJCAlignments

use of com.milaboratory.mixcr.basictypes.VDJCAlignments in project mixcr by milaboratory.

the class TargetMerger method merge.

@SuppressWarnings("unchecked")
public AlignedTarget merge(AlignedTarget targetLeft, AlignedTarget targetRight, int offset, OverlapType overlapType, int nMismatches) {
    if (offset < 0)
        return merge(targetRight, targetLeft, -offset, overlapType, nMismatches);
    final NSequenceWithQuality mergedTarget = merger.overlap(targetLeft.getTarget(), targetRight.getTarget(), offset);
    EnumMap<GeneType, VDJCHit[]> result = new EnumMap<>(GeneType.class);
    for (GeneType geneType : GeneType.VJC_REFERENCE) {
        final BatchAlignerWithBaseParameters bp = ((KGeneAlignmentParameters) alignerParameters.getGeneAlignerParameters(geneType)).getParameters();
        final VDJCHit[] leftHits = targetLeft.getAlignments().getHits(geneType);
        final VDJCHit[] rightHits = targetRight.getAlignments().getHits(geneType);
        GeneFeature alignedFeature = leftHits.length == 0 ? rightHits.length == 0 ? null : rightHits[0].getAlignedFeature() : leftHits[0].getAlignedFeature();
        Map<VDJCGeneId, HitMappingRecord> map = extractHitsMapping(targetLeft, targetRight, geneType);
        ArrayList<VDJCHit> resultingHits = new ArrayList<>();
        for (Map.Entry<VDJCGeneId, HitMappingRecord> mE : map.entrySet()) {
            final VDJCGene gene = mE.getValue().gene;
            Alignment<NucleotideSequence> mergedAl = merge(bp.getScoring(), extractBandedWidth(bp), mergedTarget.getSequence(), offset, mE.getValue().alignments[0], mE.getValue().alignments[1]);
            resultingHits.add(new VDJCHit(gene, mergedAl, alignedFeature));
        }
        Collections.sort(resultingHits);
        // final float relativeMinScore = extractRelativeMinScore(bp);
        // int threshold = (int) (resultingHits.size() > 0 ? resultingHits.get(0).getScore() * relativeMinScore : 0);
        // for (int i = resultingHits.size() - 1; i > 0; --i)
        // if (resultingHits.get(i).getScore() < threshold)
        // resultingHits.remove(i);
        result.put(geneType, resultingHits.toArray(new VDJCHit[resultingHits.size()]));
    }
    VDJCAlignments alignments = new VDJCAlignments(result, new NSequenceWithQuality[] { mergedTarget }, new SequenceHistory[] { new SequenceHistory.Merge(overlapType, targetLeft.getHistory(), targetRight.getHistory(), offset, nMismatches) }, VDJCAlignments.mergeOriginalReads(targetLeft.getAlignments(), targetRight.getAlignments()));
    AlignedTarget resultTarget = new AlignedTarget(alignments, 0);
    for (BPoint bPoint : BPoint.values()) {
        int leftPoint = targetLeft.getBPoint(bPoint);
        int rightPoint = targetRight.getBPoint(bPoint);
        if (leftPoint != -1 && rightPoint != -1)
            throw new IllegalArgumentException("Same bPoint defined in both input targets.");
        else if (leftPoint != -1)
            resultTarget = resultTarget.setBPoint(bPoint, leftPoint);
        else if (rightPoint != -1)
            resultTarget = resultTarget.setBPoint(bPoint, offset + rightPoint);
    }
    return resultTarget;
}