public void TestReadSegments()
{
var partitioned = "";
partitioned += "chr22\t1\t10\t14.00\t0\n";
partitioned += "chr22\t10\t30\t31.00\t1\n";
partitioned += "chr22\t30\t40\t6.00\t2\n";
var stringReader = new StringReader(partitioned);
Segments segments;
using (var reader = new GzipOrTextReader(stringReader))
{
segments = Segments.ReadSegments(reader);
}
Assert.Equal(segments.GetSegmentsForChromosome("chr22"), segments.AllSegments);
var confidenceInterval = segments.AllSegments.First().StartConfidenceInterval;
AssertConfidenceInterval(-5, 5, confidenceInterval);
confidenceInterval = segments.AllSegments.First().EndConfidenceInterval;
AssertConfidenceInterval(-5, 10, confidenceInterval);
confidenceInterval = segments.AllSegments[1].StartConfidenceInterval;
AssertConfidenceInterval(-5, 10, confidenceInterval);
confidenceInterval = segments.AllSegments[1].EndConfidenceInterval;
AssertConfidenceInterval(-10, 5, confidenceInterval);
confidenceInterval = segments.AllSegments.Last().StartConfidenceInterval;
AssertConfidenceInterval(-10, 5, confidenceInterval);
confidenceInterval = segments.AllSegments.Last().EndConfidenceInterval;
AssertConfidenceInterval(-5, 5, confidenceInterval);
}
public int CallVariants(string variantFrequencyFile, string inFile, string outFile, string ploidyVcfPath, string referenceFolder, string sampleName,
string truthDataPath)
{
if (!string.IsNullOrEmpty(truthDataPath))
{
_cnOracle = new CopyNumberOracle();
_cnOracle.LoadKnownCN(truthDataPath);
}
_segments = Segments.ReadSegments(_logger, new FileLocation(inFile));
_allSegments = _segments.AllSegments.ToList();
TempFolder = Path.GetDirectoryName(inFile);
if (_allSegments.Count == 0)
{
Console.WriteLine("CanvasDiploidCaller: No segments loaded; no CNV calls will be made.");
CanvasSegmentWriter.WriteSegments(outFile, _allSegments, _model?.DiploidCoverage, referenceFolder,
sampleName, null, null, QualityFilterThreshold, false, null, null);
return(0);
}
PloidyInfo ploidy = null;
if (!string.IsNullOrEmpty(ploidyVcfPath))
{
ploidy = PloidyInfo.LoadPloidyFromVcfFileNoSampleId(ploidyVcfPath);
}
// load MAF
var allelesByChromosome = CanvasIO.ReadFrequenciesWrapper(_logger, new FileLocation(variantFrequencyFile), _segments.IntervalsByChromosome);
_segments.AddAlleles(allelesByChromosome);
MeanCoverage = allelesByChromosome.SelectMany(x => x.Value).SelectMany(y => y.TotalCoverage).Average();
AggregateVariantCoverage(ref _allSegments);
// Create new models for different copy number states
InitializePloidies();
// Compute statistics on the copy number two regions
float[] diploidCounts = AggregateCounts(ref _allSegments);
_diploidCoverage = Utilities.Mean(diploidCounts);
_coverageWeightingFactor = CoverageWeighting / _diploidCoverage;
// new coverage model
_model = new CoverageModel {
DiploidCoverage = _diploidCoverage
};
List <SegmentInfo> segments = new List <SegmentInfo>();
foreach (CanvasSegment segment in _allSegments)
{
SegmentInfo info = new SegmentInfo {
Segment = segment
};
List <double> mafs = new List <double>();
foreach (float value in segment.Balleles.Frequencies)
{
mafs.Add(value > 0.5 ? 1 - value : value);
}
if (mafs.Count > 0)
{
info.Maf = Utilities.Median(mafs);
}
else
{
info.Maf = -1;
}
info.Coverage = Utilities.Median(segment.Counts);
info.Weight = _allSegments.Count > 100 ? segment.Length : segment.BinCount;
segments.Add(info);
}
AssignPloidyCallsDistance(_model);
CanvasSegment.AssignQualityScores(_allSegments, CanvasSegment.QScoreMethod.LogisticGermline, _germlineScoreParameters);
// Merge neighboring segments that got the same copy number call.
// merging segments requires quality scores so we do it after quality scores have been assigned
var mergedSegments = CanvasSegment.MergeSegments(_allSegments);
// recalculating qscores after merging segments improves performance!
CanvasSegment.AssignQualityScores(mergedSegments, CanvasSegment.QScoreMethod.LogisticGermline, _germlineScoreParameters);
CanvasSegment.SetFilterForSegments(QualityFilterThreshold, mergedSegments, CanvasFilter.SegmentSizeCutoff);
List <string> extraHeaders = new List <string>();
var coverageOutputPath = SingleSampleCallset.GetCoverageAndVariantFrequencyOutputPath(outFile);
CanvasSegment.WriteCoveragePlotData(mergedSegments, _model.DiploidCoverage, ploidy, coverageOutputPath, referenceFolder);
if (_cnOracle != null)
{
GenerateReportVersusKnownCopyNumber();
}
if (!string.IsNullOrEmpty(ploidy?.HeaderLine))
{
extraHeaders.Add(ploidy.HeaderLine);
}
CanvasSegmentWriter.WriteSegments(outFile, mergedSegments, _model.DiploidCoverage, referenceFolder, sampleName,
extraHeaders, ploidy, QualityFilterThreshold, false, null, null);
return(0);
}
internal int CallVariants(List <string> variantFrequencyFiles, List <string> segmentFiles,
IFileLocation outVcfFile, string ploidyBedPath, string referenceFolder, List <string> sampleNames, string commonCnvsBedPath, List <SampleType> sampleTypes)
{
// load files
// initialize data structures and classes
var fileCounter = 0;
var samplesInfo = new SampleMap <SampleMetrics>();
var sampleSegments = new SampleMap <Segments>();
var copyNumberModels = new SampleMap <ICopyNumberModel>();
var variantFrequencyFilesSampleList = new SampleMap <string>();
var kinships = new SampleMap <SampleType>();
foreach (string sampleName in sampleNames)
{
var sampleId = new SampleId(sampleName);
var segment = Segments.ReadSegments(_logger, new FileLocation(segmentFiles[fileCounter]));
segment.AddAlleles(CanvasIO.ReadFrequenciesWrapper(_logger, new FileLocation(variantFrequencyFiles[fileCounter]), segment.IntervalsByChromosome));
sampleSegments.Add(sampleId, segment);
var sampleInfo = SampleMetrics.GetSampleInfo(segment.AllSegments, ploidyBedPath, _callerParameters.NumberOfTrimmedBins, sampleId);
var copyNumberModel = _copyNumberModelFactory.CreateModel(_callerParameters.MaximumCopyNumber, sampleInfo.MaxCoverage, sampleInfo.MeanCoverage, sampleInfo.MeanMafCoverage);
samplesInfo.Add(sampleId, sampleInfo);
copyNumberModels.Add(sampleId, copyNumberModel);
variantFrequencyFilesSampleList.Add(sampleId, variantFrequencyFiles[fileCounter]);
kinships.Add(sampleId, sampleTypes[fileCounter]);
fileCounter++;
}
var segmentSetsFromCommonCnvs = CreateSegmentSetsFromCommonCnvs(variantFrequencyFilesSampleList,
_callerParameters.MinAlleleCountsThreshold, commonCnvsBedPath, sampleSegments);
var segmentsForVariantCalling = GetHighestLikelihoodSegments(segmentSetsFromCommonCnvs, samplesInfo, copyNumberModels).ToList();
PedigreeInfo pedigreeInfo = PedigreeInfo.GetPedigreeInfo(kinships, _callerParameters);
Parallel.ForEach(
segmentsForVariantCalling,
new ParallelOptions
{
MaxDegreeOfParallelism = Math.Min(Environment.ProcessorCount, _callerParameters.MaxCoreNumber)
},
segments => _variantCaller.CallVariant(segments, samplesInfo, copyNumberModels, pedigreeInfo)
);
var variantCalledSegments = new SampleMap <List <CanvasSegment> >();
foreach (var key in samplesInfo.SampleIds)
{
variantCalledSegments.Add(key, segmentsForVariantCalling.Select(segment => segment[key]).ToList());
}
var mergedVariantCalledSegments = MergeSegments(variantCalledSegments, _callerParameters.MinimumCallSize, _qualityFilterThreshold);
FilterExcessivelyShortSegments(mergedVariantCalledSegments);
var outputFolder = outVcfFile.Directory;
foreach (var sampleId in samplesInfo.SampleIds)
{
var coverageOutputPath = SingleSampleCallset.GetCoverageAndVariantFrequencyOutput(outputFolder,
sampleId.ToString());
CanvasSegment.WriteCoveragePlotData(mergedVariantCalledSegments[sampleId], samplesInfo[sampleId].MeanCoverage,
samplesInfo[sampleId].Ploidy, coverageOutputPath, referenceFolder);
}
bool isPedigreeInfoSupplied = pedigreeInfo != null && pedigreeInfo.HasFullPedigree();
var denovoQualityThreshold = isPedigreeInfoSupplied ? (int?)_deNovoQualityFilterThreshold : null;
var ploidies = samplesInfo.Select(info => info.Value.Ploidy).ToList();
var diploidCoverage = samplesInfo.Select(info => info.Value.MeanCoverage).ToList();
var names = samplesInfo.SampleIds.Select(id => id.ToString()).ToList();
CanvasSegmentWriter.WriteMultiSampleSegments(outVcfFile.FullName, mergedVariantCalledSegments, diploidCoverage, referenceFolder, names,
null, ploidies, _qualityFilterThreshold, denovoQualityThreshold, CanvasFilter.SegmentSizeCutoff, isPedigreeInfoSupplied);
foreach (var sampleId in samplesInfo.SampleIds)
{
var outputVcfPath = SingleSampleCallset.GetVcfOutput(outputFolder, sampleId.ToString());
var sampleMetrics = samplesInfo[sampleId];
var segments = mergedVariantCalledSegments[sampleId];
CanvasSegmentWriter.WriteSegments(outputVcfPath.FullName, segments,
sampleMetrics.MeanCoverage, referenceFolder, sampleId.ToString(), null,
sampleMetrics.Ploidy, _qualityFilterThreshold, isPedigreeInfoSupplied, denovoQualityThreshold, null);
var visualizationTemp = outputFolder.CreateSubdirectory($"VisualizationTemp{sampleId}");
var normalizationFactor = NormalizationCalculator.ComputeNormalizationFactor(segments);
var bigWig = _coverageBigWigWriter.Write(segments, visualizationTemp, normalizationFactor);
bigWig?.MoveTo(SingleSampleCallset.GetCoverageBigWig(outputFolder, sampleId.ToString()));
var copyNumberBedGraph = SingleSampleCallset.GetCopyNumberBedGraph(outputFolder, sampleId.ToString());
_copyNumberBedGraphWriter.Write(segments, sampleMetrics.Ploidy, copyNumberBedGraph);
var partitionBedgraphHeader = "track type=bedGraph visibility=full autoScale=on graphType=points";
var originalSegments = sampleSegments[sampleId];
_partitionCoverageBedGraphWriter.Write(originalSegments.AllSegments, SingleSampleCallset.GetPartitionBedGraph(outputFolder, sampleId.ToString()), normalizationFactor, partitionBedgraphHeader);
}
return(0);
}