using Test using BioGenerics using FormatSpecimens using GenomicFeatures using XAM import BioAlignments: Alignment, AlignmentAnchor, OP_START, OP_MATCH, OP_DELETE import BGZFStreams: BGZFStream import BioGenerics.Exceptions: MissingFieldException import BioSequences: @dna_str, @aa_str # Generate a random range within `range`. function randrange(range) x = rand(range) y = rand(range) if x < y return x:y else return y:x end end @testset "SAM" begin samdir = path_of_format("SAM") @testset "MetaInfo" begin metainfo = SAM.MetaInfo() @test !isfilled(metainfo) @test occursin("not filled", repr(metainfo)) metainfo = SAM.MetaInfo("CO", "some comment (parens)") @test isfilled(metainfo) @test string(metainfo) == "@CO\tsome comment (parens)" @test occursin("CO", repr(metainfo)) @test SAM.tag(metainfo) == "CO" @test SAM.value(metainfo) == "some comment (parens)" @test_throws ArgumentError keys(metainfo) @test_throws ArgumentError values(metainfo) metainfo = SAM.MetaInfo("HD", ["VN" => "1.0", "SO" => "coordinate"]) @test isfilled(metainfo) @test string(metainfo) == "@HD\tVN:1.0\tSO:coordinate" @test occursin("HD", repr(metainfo)) @test SAM.tag(metainfo) == "HD" @test SAM.value(metainfo) == "VN:1.0\tSO:coordinate" @test keys(metainfo) == ["VN", "SO"] @test values(metainfo) == ["1.0", "coordinate"] @test SAM.keyvalues(metainfo) == ["VN" => "1.0", "SO" => "coordinate"] @test haskey(metainfo, "VN") @test haskey(metainfo, "SO") @test !haskey(metainfo, "GO") @test metainfo["VN"] == "1.0" @test metainfo["SO"] == "coordinate" @test_throws KeyError metainfo["GO"] end @testset "Header" begin header = SAM.Header() @test isempty(header) push!(header, SAM.MetaInfo("@HD\tVN:1.0\tSO:coordinate")) @test !isempty(header) @test length(header) == 1 push!(header, SAM.MetaInfo("@CO\tsome comment")) @test length(header) == 2 @test isa(collect(header), Vector{SAM.MetaInfo}) end @testset "Record" begin record = SAM.Record() @test !isfilled(record) @test !SAM.ismapped(record) @test repr(record) == "XAM.SAM.Record: " @test_throws ArgumentError SAM.flag(record) record = SAM.Record("r001\t99\tchr1\t7\t30\t8M2I4M1D3M\t=\t37\t39\tTTAGATAAAGGATACTG\t*") @test isfilled(record) @test occursin(r"^XAM.SAM.Record:\n", repr(record)) @test SAM.ismapped(record) @test SAM.isprimary(record) @test SAM.hastempname(record) @test SAM.tempname(record) == "r001" @test SAM.hasflag(record) @test SAM.flag(record) === UInt16(99) @test SAM.hasrefname(record) @test SAM.refname(record) == "chr1" @test SAM.hasposition(record) @test SAM.position(record) === 7 @test SAM.hasmappingquality(record) @test SAM.mappingquality(record) === UInt8(30) @test SAM.hascigar(record) @test SAM.cigar(record) == "8M2I4M1D3M" @test SAM.hasnextrefname(record) @test SAM.nextrefname(record) == "=" @test SAM.hasnextposition(record) @test SAM.nextposition(record) === 37 @test SAM.hastemplength(record) @test SAM.templength(record) === 39 @test SAM.hassequence(record) @test SAM.sequence(record) == dna"TTAGATAAAGGATACTG" @test !SAM.hasquality(record) @test_throws MissingFieldException SAM.quality(record) end @testset "Reader" begin reader = open(SAM.Reader, joinpath(samdir, "ce#1.sam")) @test isa(reader, SAM.Reader) @test eltype(reader) === SAM.Record # header h = header(reader) @test string.(findall(header(reader), "SQ")) == ["@SQ\tSN:CHROMOSOME_I\tLN:1009800"] # first record record = SAM.Record() read!(reader, record) @test SAM.ismapped(record) @test SAM.refname(record) == "CHROMOSOME_I" @test SAM.position(record) == leftposition(record) == 2 @test SAM.rightposition(record) == rightposition(record) == 102 @test SAM.tempname(record) == seqname(record) == "SRR065390.14978392" @test SAM.sequence(record) == sequence(record) == dna"CCTAGCCCTAACCCTAACCCTAACCCTAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAA" @test SAM.sequence(String, record) == "CCTAGCCCTAACCCTAACCCTAACCCTAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAA" @test SAM.seqlength(record) == 100 @test SAM.quality(record) == (b"#############################@B?8B?BA@@DDBCDDCBC@CDCDCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCC" .- 33) @test SAM.quality(String, record) == "#############################@B?8B?BA@@DDBCDDCBC@CDCDCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCC" @test SAM.flag(record) == 16 @test SAM.cigar(record) == "27M1D73M" @test SAM.alignment(record) == Alignment([ AlignmentAnchor( 0, 1, OP_START), AlignmentAnchor( 27, 28, OP_MATCH), AlignmentAnchor( 27, 29, OP_DELETE), AlignmentAnchor(100, 102, OP_MATCH)]) @test record["XG"] == 1 @test record["XM"] == 5 @test record["XN"] == 0 @test record["XO"] == 1 @test record["AS"] == -18 @test record["XS"] == -18 @test record["YT"] == "UU" @test eof(reader) close(reader) # rightposition (also implicitly alignlength) records = collect(open(SAM.Reader, joinpath(samdir, "ce#5b.sam"))) @test SAM.rightposition(records[6]) == rightposition(records[6]) == 83 # iterator @test length(collect(open(SAM.Reader, joinpath(samdir, "ce#1.sam")))) == 1 @test length(collect(open(SAM.Reader, joinpath(samdir, "ce#2.sam")))) == 2 # IOStream @test length(collect(SAM.Reader(open(joinpath(samdir, "ce#1.sam"))))) == 1 @test length(collect(SAM.Reader(open(joinpath(samdir, "ce#2.sam"))))) == 2 end @testset "Round trip" begin function compare_records(xs, ys) if length(xs) != length(ys) return false end for (x, y) in zip(xs, ys) if x.data[x.filled] != y.data[y.filled] return false end end return true end for specimen in list_valid_specimens("SAM") filepath = joinpath(samdir, filename(specimen)) mktemp() do path, io # copy reader = open(SAM.Reader, filepath) writer = SAM.Writer(io, header(reader)) records = SAM.Record[] for record in reader push!(records, record) write(writer, record) end close(reader) close(writer) @test compare_records(open(collect, SAM.Reader, path), records) end end end end @testset "BAM" begin bamdir = path_of_format("BAM") @testset "AuxData" begin auxdata = BAM.AuxData(UInt8[]) @test isempty(auxdata) buf = IOBuffer() write(buf, "NM", UInt8('s'), Int16(1)) auxdata = BAM.AuxData(take!(buf)) @test length(auxdata) == 1 @test auxdata["NM"] === Int16(1) @test collect(auxdata) == ["NM" => Int16(1)] buf = IOBuffer() write(buf, "AS", UInt8('c'), Int8(-18)) write(buf, "NM", UInt8('s'), Int16(1)) write(buf, "XA", UInt8('f'), Float32(3.14)) write(buf, "XB", UInt8('Z'), "some text\0") write(buf, "XC", UInt8('B'), UInt8('i'), Int32(3), Int32[10, -5, 8]) auxdata = BAM.AuxData(take!(buf)) @test length(auxdata) == 5 @test auxdata["AS"] === Int8(-18) @test auxdata["NM"] === Int16(1) @test auxdata["XA"] === Float32(3.14) @test auxdata["XB"] == "some text" @test auxdata["XC"] == Int32[10, -5, 8] @test convert(Dict{String,Any}, auxdata) == Dict( "AS" => Int8(-18), "NM" => Int16(1), "XA" => Float32(3.14), "XB" => "some text", "XC" => Int32[10, -5, 8]) end @testset "Record" begin record = BAM.Record() @test !isfilled(record) @test repr(record) == "XAM.BAM.Record: " @test_throws ArgumentError BAM.flag(record) end @testset "Reader" begin reader = open(BAM.Reader, joinpath(bamdir, "ce#1.bam")) @test isa(reader, BAM.Reader) @test eltype(reader) === BAM.Record @test startswith(repr(reader), "XAM.BAM.Reader{IOStream}:") # header h = header(reader) @test isa(h, SAM.Header) # first record record = BAM.Record() read!(reader, record) @test BAM.ismapped(record) @test BAM.isprimary(record) @test ! BAM.ispositivestrand(record) @test BAM.refname(record) == "CHROMOSOME_I" @test BAM.refid(record) === 1 @test BAM.hasnextrefid(record) @test BAM.nextrefid(record) === 0 @test BAM.hasposition(record) === hasleftposition(record) === true @test BAM.position(record) === leftposition(record) === 2 @test BAM.hasnextposition(record) @test BAM.nextposition(record) === 0 @test rightposition(record) == 102 @test BAM.hastempname(record) === hasseqname(record) === true @test BAM.tempname(record) == seqname(record) == "SRR065390.14978392" @test BAM.hassequence(record) === hassequence(record) === true @test BAM.sequence(record) == sequence(record) == dna""" CCTAGCCCTAACCCTAACCCTAACCCTAGCCTAAGCCTAAGCCTAAGCCT AAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAAGCCTAA """ @test BAM.seqlength(record) === 100 @test BAM.hasquality(record) @test eltype(BAM.quality(record)) == UInt8 @test BAM.quality(record) == [Int(x) - 33 for x in "#############################@B?8B?BA@@DDBCDDCBC@CDCDCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCC"] @test BAM.flag(record) === UInt16(16) @test BAM.cigar(record) == "27M1D73M" @test BAM.alignment(record) == Alignment([ AlignmentAnchor( 0, 1, OP_START), AlignmentAnchor( 27, 28, OP_MATCH), AlignmentAnchor( 27, 29, OP_DELETE), AlignmentAnchor(100, 102, OP_MATCH)]) @test record["XG"] == 1 @test record["XM"] == 5 @test record["XN"] == 0 @test record["XO"] == 1 @test record["AS"] == -18 @test record["XS"] == -18 @test record["YT"] == "UU" @test keys(record) == ["XG","XM","XN","XO","AS","XS","YT"] @test values(record) == [1, 5, 0, 1, -18, -18, "UU"] @test eof(reader) close(reader) # Test conversion from byte array to record dsize = BAM.data_size(record) array = Vector{UInt8}(undef, BAM.FIXED_FIELDS_BYTES + dsize) GC.@preserve array record begin ptr = Ptr{UInt8}(pointer_from_objref(record)) unsafe_copyto!(pointer(array), ptr, BAM.FIXED_FIELDS_BYTES) unsafe_copyto!(array, BAM.FIXED_FIELDS_BYTES + 1, record.data, 1, dsize) end new_record = convert(BAM.Record, array) @test record.bin_mq_nl == new_record.bin_mq_nl @test record.block_size == new_record.block_size @test record.flag_nc == new_record.flag_nc @test record.l_seq == new_record.l_seq @test record.next_refid == new_record.next_refid @test record.next_pos == new_record.next_pos @test record.refid == new_record.refid @test record.pos == new_record.pos @test record.tlen == new_record.tlen @test record.data == new_record.data # rightposition (also implicitly alignlength) records = collect(open(BAM.Reader, joinpath(bamdir, "ce#5b.bam"))) @test BAM.rightposition(records[6]) == rightposition(records[6]) == 83 # iterator @test length(collect(open(BAM.Reader, joinpath(bamdir, "ce#1.bam")))) == 1 @test length(collect(open(BAM.Reader, joinpath(bamdir, "ce#2.bam")))) == 2 # IOStream @test length(collect(BAM.Reader(open(joinpath(bamdir, "ce#1.bam"))))) == 1 @test length(collect(BAM.Reader(open(joinpath(bamdir, "ce#2.bam"))))) == 2 end @testset "Read long CIGARs" begin function get_cigar_lens(rec::BAM.Record) cigar_ops, cigar_n = BAM.cigar_rle(rec) field_ops, field_n = BAM.cigar_rle(rec, false) cigar_l = length(cigar_ops) field_l = length(field_ops) return cigar_l, field_l end function check_cigar_vs_field(rec::BAM.Record) cigar = BAM.cigar(rec) field = BAM.cigar(rec, false) cigar_l, field_l = get_cigar_lens(rec) return cigar != field && cigar_l != field_l end function check_cigar_lens(rec::BAM.Record, field_len, cigar_len) cigar_l, field_l = get_cigar_lens(rec) return cigar_l == cigar_len && field_l == field_len end reader = open(BAM.Reader, joinpath(bamdir, "cigar-64k.bam")) rec = BAM.Record() read!(reader, rec) @test !check_cigar_vs_field(rec) read!(reader, rec) @test check_cigar_vs_field(rec) @test check_cigar_lens(rec, 2, 72091) end function compare_records(xs, ys) if length(xs) != length(ys) return false end for (x, y) in zip(xs, ys) if !( x.block_size == y.block_size && x.refid == y.refid && x.pos == y.pos && x.bin_mq_nl == y.bin_mq_nl && x.flag_nc == y.flag_nc && x.l_seq == y.l_seq && x.next_refid == y.next_refid && x.next_pos == y.next_pos && x.tlen == y.tlen && x.data[1:BAM.data_size(x)] == y.data[1:BAM.data_size(y)]) return false end end return true end @testset "Round trip" begin for specimen in list_valid_specimens("BAM") filepath = joinpath(bamdir, filename(specimen)) mktemp() do path, _ # copy if hastags(specimen) && in("bai", tags(specimen)) reader = open(BAM.Reader, filepath, index=filepath * ".bai") else reader = open(BAM.Reader, filepath) end writer = BAM.Writer( BGZFStream(path, "w"), BAM.header(reader, fillSQ=isempty(findall(header(reader), "SQ")))) records = BAM.Record[] for record in reader push!(records, record) write(writer, record) end close(reader) close(writer) @test compare_records(open(collect, BAM.Reader, path), records) end end end @testset "Random access" begin filepath = joinpath(bamdir, "GSE25840_GSM424320_GM06985_gencode_spliced.head.bam") reader = open(BAM.Reader, filepath, index=filepath * ".bai") @test isa(eachoverlap(reader, "chr1", 1:100), BAM.OverlapIterator) @test isa(eachoverlap(reader, GenomicFeatures.Interval("chr1", 1, 100)), BAM.OverlapIterator) # expected values are counted using samtools for (refname, interval, expected) in [ ("chr1", 1_000:10000, 21), ("chr1", 8_000:10000, 20), ("chr1", 766_000:800_000, 142), ("chr1", 786_000:800_000, 1), ("chr1", 796_000:800_000, 0)] intsect = eachoverlap(reader, refname, interval) @test eltype(intsect) == BAM.Record @test count(_ -> true, intsect) == expected # check that the intersection iterator is stateless @test count(_ -> true, intsect) == expected end # randomized tests for n in 1:50 refindex = 1 refname = "chr1" range = randrange(1:1_000_000) seekstart(reader) # linear scan expected = filter(collect(reader)) do record BAM.compare_intervals(record, (refindex, range)) == 0 end # indexed scan actual = collect(eachoverlap(reader, refname, range)) @test compare_records(actual, expected) end close(reader) filepath = joinpath(bamdir, "R_12h_D06.uniq.q40.bam") reader = open(BAM.Reader, filepath, index=filepath * ".bai") @test isempty(collect(eachoverlap(reader, "chr19", 5823708:5846478))) close(reader) end end