'''Tools for working with files in the samtools pileup -c format.''' import collections import pysam PileupSubstitution = collections.namedtuple("PileupSubstitution", " ".join(( "chromosome", "pos", "reference_base", "genotype", "consensus_quality", "snp_quality", "mapping_quality", "coverage", "read_bases", "base_qualities"))) PileupIndel = collections.namedtuple("PileupIndel", " ".join(( "chromosome", "pos", "reference_base", "genotype", "consensus_quality", "snp_quality", "mapping_quality", "coverage", "first_allele", "second_allele", "reads_first", "reads_second", "reads_diff"))) def iterate(infile): '''iterate over ``samtools pileup -c`` formatted file. *infile* can be any iterator over a lines. The function yields named tuples of the type :class:`pysam.Pileup.PileupSubstitution` or :class:`pysam.Pileup.PileupIndel`. .. note:: The parser converts to 0-based coordinates ''' conv_subst = (str, lambda x: int(x) - 1, str, str, int, int, int, int, str, str) conv_indel = (str, lambda x: int(x) - 1, str, str, int, int, int, int, str, str, int, int, int) for line in infile: d = line[:-1].split() if d[2] == "*": try: yield PileupIndel(*[x(y) for x, y in zip(conv_indel, d)]) except TypeError: raise pysam.SamtoolsError("parsing error in line: `%s`" % line) else: try: yield PileupSubstitution(*[x(y) for x, y in zip(conv_subst, d)]) except TypeError: raise pysam.SamtoolsError("parsing error in line: `%s`" % line) ENCODE_GENOTYPE = { 'A': 'A', 'C': 'C', 'G': 'G', 'T': 'T', 'AA': 'A', 'CC': 'C', 'GG': 'G', 'TT': 'T', 'UU': 'U', 'AG': 'r', 'GA': 'R', 'CT': 'y', 'TC': 'Y', 'AC': 'm', 'CA': 'M', 'GT': 'k', 'TG': 'K', 'CG': 's', 'GC': 'S', 'AT': 'w', 'TA': 'W', } DECODE_GENOTYPE = { 'A': 'AA', 'C': 'CC', 'G': 'GG', 'T': 'TT', 'r': 'AG', 'R': 'AG', 'y': 'CT', 'Y': 'CT', 'm': 'AC', 'M': 'AC', 'k': 'GT', 'K': 'GT', 's': 'CG', 'S': 'CG', 'w': 'AT', 'W': 'AT', } # ------------------------------------------------------------ def encodeGenotype(code): '''encode genotypes like GG, GA into a one-letter code. The returned code is lower case if code[0] < code[1], otherwise it is uppercase. ''' return ENCODE_GENOTYPE[code.upper()] def decodeGenotype(code): '''decode single letter genotypes like m, M into two letters. This is the reverse operation to :meth:`encodeGenotype`. ''' return DECODE_GENOTYPE[code] def translateIndelGenotypeFromVCF(vcf_genotypes, ref): '''translate indel from vcf to pileup format.''' # indels def getPrefix(s1, s2): '''get common prefix of strings s1 and s2.''' n = min(len(s1), len(s2)) for x in range(n): if s1[x] != s2[x]: return s1[:x] return s1[:n] def getSuffix(s1, s2): '''get common sufix of strings s1 and s2.''' n = min(len(s1), len(s2)) if s1[-1] != s2[-1]: return "" for x in range(-2, -n - 1, -1): if s1[x] != s2[x]: return s1[x + 1:] return s1[-n:] def getGenotype(variant, ref): if variant == ref: return "*", 0 if len(ref) > len(variant): # is a deletion if ref.startswith(variant): return "-%s" % ref[len(variant):], len(variant) - 1 elif ref.endswith(variant): return "-%s" % ref[:-len(variant)], -1 else: prefix = getPrefix(ref, variant) suffix = getSuffix(ref, variant) shared = len(prefix) + len(suffix) - len(variant) # print "-", prefix, suffix, ref, variant, shared, len(prefix), len(suffix), len(ref) if shared < 0: raise ValueError() return "-%s" % ref[len(prefix):-(len(suffix) - shared)], len(prefix) - 1 elif len(ref) < len(variant): # is an insertion if variant.startswith(ref): return "+%s" % variant[len(ref):], len(ref) - 1 elif variant.endswith(ref): return "+%s" % variant[:len(ref)], 0 else: prefix = getPrefix(ref, variant) suffix = getSuffix(ref, variant) shared = len(prefix) + len(suffix) - len(ref) if shared < 0: raise ValueError() return "+%s" % variant[len(prefix):-(len(suffix) - shared)], len(prefix) else: assert 0, "snp?" # in pileup, the position refers to the base # after the coordinate, hence subtract 1 # pos -= 1 genotypes, offsets = [], [] is_error = True for variant in vcf_genotypes: try: g, offset = getGenotype(variant, ref) except ValueError: break genotypes.append(g) if g != "*": offsets.append(offset) else: is_error = False if is_error: raise ValueError() assert len(set(offsets)) == 1, "multiple offsets for indel" offset = offsets[0] genotypes = "/".join(genotypes) return genotypes, offset def vcf2pileup(vcf, sample): '''convert vcf record to pileup record.''' chromosome = vcf.contig pos = vcf.pos reference = vcf.ref allelles = [reference] + vcf.alt data = vcf[sample] # get genotype genotypes = data["GT"] if len(genotypes) > 1: raise ValueError("only single genotype per position, %s" % (str(vcf))) genotypes = genotypes[0] # not a variant if genotypes[0] == ".": return None genotypes = [allelles[int(x)] for x in genotypes if x != "/"] # snp_quality is "genotype quality" snp_quality = consensus_quality = data.get("GQ", [0])[0] mapping_quality = vcf.info.get("MQ", [0])[0] coverage = data.get("DP", 0) if len(reference) > 1 or max([len(x) for x in vcf.alt]) > 1: # indel genotype, offset = translateIndelGenotypeFromVCF(genotypes, reference) return PileupIndel(chromosome, pos + offset, "*", genotype, consensus_quality, snp_quality, mapping_quality, coverage, genotype, "<" * len(genotype), 0, 0, 0) else: genotype = encodeGenotype("".join(genotypes)) read_bases = "" base_qualities = "" return PileupSubstitution(chromosome, pos, reference, genotype, consensus_quality, snp_quality, mapping_quality, coverage, read_bases, base_qualities) def iterate_from_vcf(infile, sample): '''iterate over a vcf-formatted file. *infile* can be any iterator over a lines. The function yields named tuples of the type :class:`pysam.Pileup.PileupSubstitution` or :class:`pysam.Pileup.PileupIndel`. Positions without a snp will be skipped. This method is wasteful and written to support same legacy code that expects samtools pileup output. Better use the vcf parser directly. ''' vcf = pysam.VCF() vcf.connect(infile) if sample not in vcf.getsamples(): raise KeyError("sample %s not vcf file") for row in vcf.fetch(): result = vcf2pileup(row, sample) if result: yield result