def main():
""" getting the git hash in python: http://stackoverflow.com/a/14989911/1735942"""
label = subprocess.check_output(["git", "rev-parse", "HEAD"])
today=datetime.datetime.today()
datestr=today.strftime("20%y%m%d")
usage = "usage: %prog [options] my.bam "
parser = OptionParser(usage)
parser.add_option("--bed", type="string", dest="bedfile", help="bed file with coordinates")
parser.add_option("--tbf", type="string", dest="tbfile", help=" *.2bit file of the reference sequence")
parser.add_option("--ped", type="string", dest="pedfile", help= " pedfile of the samples you are analyzing")
parser.add_option("--min-nonref-count", dest="minAlt", default=2, help="minimum observation of nonref allele for genotype inference (default 2)")
(options, args)=parser.parse_args()
commandline=";".join(sys.argv)
bamfile=args[0]
bamfilebasename=return_file_basename(bamfile)
vcfoutput=".".join(['pgmsnp',bamfilebasename, datestr, 'vcf'])
vcfh=open(vcfoutput,'w')
ALPHABET=['AA', 'AC', 'AG', 'AT', 'CC', 'CG', 'CT', 'GG', 'GT', 'TT']
twobit=bx.seq.twobit.TwoBitFile( open( options.tbfile ) )
bedobj=Bedfile(options.bedfile)
pybamfile=pysam.Samfile( bamfile, "rb" )
pedfile=Pedfile(options.pedfile)
pedfile.parsePedfile()
samplenames=pedfile.returnIndivids()
samplestring="\t".join(samplenames)
#vcf metainfolines
vcf_metalines=[]
vcf_metalines.append ( "##fileformat=VCFv4.1")
vcf_metalines.append( "##fileDate="+datestr )
vcf_metalines.append("##testGeneticFactor="+commandline)
vcf_metalines.append("##version="+label.strip())
vcf_metalines.append("##reference="+options.tbfile)
vcf_metalines.append("##pedfile="+options.pedfile)
vcf_metalines.append("##bedfile="+options.bedfile)
vcf_metalines.append("##bamfile="+bamfile)
vcf_metalines.append("##INFO=<ID=NS,Number=1,Type=Integer,Description=\"Number of Samples With Data\">")
vcf_metalines.append( "##INFO=<ID=DP,Number=1,Type=Integer,Description=\"Total Depth\">" )
vcf_metalines.append( "##FORMAT=<ID=GT,Number=1,Type=String,Description=\"Genotype\">" )
vcf_metalines.append( "##FORMAT=<ID=GQ,Number=1,Type=Integer,Description=\"Genotype Quality\">" )
vcf_metalines.append( "##FORMAT=<ID=DP,Number=1,Type=Integer,Description=\"Read Depth\">" )
vcf_column_headers=["#CHROM", "POS", "ID", "REF", "ALT", "QUAL", "FILTER", "INFO", "FORMAT",samplestring]
vcf_metalines.append( "\t".join(vcf_column_headers))
vcf_metaline_output="\n".join(vcf_metalines)
vcfh.write(vcf_metaline_output+"\n")
#print vcf_metaline_output
#print samplenames
#return the complete enumeration of all possible offspring genotype priors
punnetValues=returnPunnetValues(4)
# Pfactor gives us a pileup iterator
Pfactory=PileupFactory(pybamfile,bedobj)
##CHROM", "POS", "ID", "REF", "ALT", "QUAL", "FILTER", "INFO", "FORMAT",samplestring
for pileup_data_obj in Pfactory.yieldPileupData():
refDepth=defaultdict(lambda: defaultdict(int))
altDepth=defaultdict(lambda: defaultdict(int))
pileup_column_pos=pileup_data_obj.getPileupColumnPos()
pileup_column_chrom=pileup_data_obj.getChrom()
refsequence=twobit[pileup_column_chrom][pileup_column_pos:pileup_column_pos+1] #this is the reference base
refDepth=defaultdict(lambda: defaultdict(int))
altDepth=defaultdict(lambda: defaultdict(int))
skipsite=False
#print 'Pileup position: ', pileup_data_obj, "\t".join( [pileup_column_chrom, str(pileup_column_pos), refsequence])
#print
qual="."
filter='.'
siteDP="DP="+str(pileup_data_obj.getPileupDepth())
altDP=0
sampleDepth=defaultdict(int)
#lets make our genetic network here:
pgmNetwork=PgmNetworkFactory(options.pedfile,pileup_column_chrom,pileup_column_pos, refsequence,punnetValues)
totalSize=pgmNetwork.returnTotalSize()
#print "totalSize: ", totalSize
observedSamples=[]
sampleNames=set(pgmNetwork.getSampleNames())
#print "pgmNetwork factor list: "
#pgmNetwork.printFactorList()
pgmFactorList=pgmNetwork.getFactorList()
#print "++++"
for (sample, pileup_sample_data) in pileup_data_obj.yieldSamplePileupData():
sampleDepth[sample]=len(pileup_sample_data)
observedSamples.append(sample)
sample_idx=pgmNetwork.getSampleNamePedIndex(sample)
for data in pileup_sample_data:
if data.basecall != refsequence: altDepth[data.sample][data.basecall]+=1
if data.basecall == refsequence: refDepth[data.sample][data.basecall]+=1
value=calculateGLL(pileup_sample_data)
pgmFactorList[sample_idx + totalSize].setVal(value)
#we initially get the log-likelihood, but go back to probablity space first
pgmFactorList[sample_idx + totalSize] = ExpFactor( pgmFactorList[sample_idx + totalSize] )
#print pgmFactorList[sample_idx + totalSize]
#GLFactor = Factor( [readVar, genoVar], [1,10], [], 'read_phenotype | genotype ')
#gPrior=LogFactor( returnGenotypePriorFounderFactor(sequence,['A','C','G','T'], genoVar) )
#print
if sum(chain.from_iterable(d.itervalues() for d in altDepth.itervalues())) < options.minAlt:
#print pileup_data_obj.getPileupColumnPos(), altDP
continue
observedSamples=set(observedSamples)
unobservedSamples=sampleNames-observedSamples
unobservedIdxs=[ pgmNetwork.getSampleNamePedIndex(sample) for sample in unobservedSamples ]
#for samples lacking read data, delete them from the list of
for idx in unobservedIdxs:
#del pgmFactorList[idx + totalSize]
value=calculateNoObservationsGL()
#pdb.set_trace()
pgmFactorList[idx + totalSize].setVal(value)
#print pgmFactorList
#for f in pgmFactorList:
# print f
#continue
#print "====="
siteNS="NS="+str(len(observedSamples))
infoString=";".join([siteNS, siteDP])
#for f in pgmFactorList:
# print f
# print
#pdb.set_trace()
#cTree=CreatePrunedInitCtree(pgmFactorList)
#print 'cTree constructed, pruned, and initalized potential'
#G=nx.from_numpy_matrix( cTree.getEdges() )
#nx.draw_shell(G)
#plt.show()
#print cTree
#get the max marginal factors
MAXMARGINALS=ComputeExactMarginalsBP(pgmFactorList, [], 1)
#MARGINALS= ComputeExactMarginalsBP( pgmFactorList)
#this log normalizes the data
for m in MAXMARGINALS:
#print m.getVal()
m.setVal( np.log( lognormalize(m.getVal() ) ) )
#print np.sum( lognormalize(m.getVal() ) )
#print'lognormalize: ', lognormalize(m.getVal() )
#print m.getVal()
#print
# print m.getVal()
# print
#print "==="
#get the max value of each factor
MAXvalues=[ max(m.getVal().tolist() ) for m in MAXMARGINALS ]
#MAXvalues_phred = [ PhredScore(x) for x in MAXvalues ]
""" phred scale the error prob. so we take the 1-posterior, and phred scale it
Note we have expoentiate the values then take the complement, then phred scale """
phred_gq= [ round(PhredScore(x),2) for x in (1-np.exp(MAXvalues[0:totalSize])) ]
#MAXvalues_str= [str(x) for x in MAXvalues]
phred_str=[str(x) for x in phred_gq]
#print "maxvalues: ", MAXvalues[0:totalSize]
#this is the decoding, where we get teh assignment of the variable
# that has the max marginoal value
MAPAssignment=MaxDecoding( MAXMARGINALS )
#print "MAPAssignment: ", MAPAssignment[0:3]
#print totalSize
#for idx in range(totalSize):
# print ALPHABET[ MAPAssignment[idx] ]
#print MAPAssignment
#we convert from variable assignment in the factor to genotype space
sampleNames=pgmNetwork.returnGenotypeFactorIdxSampleNames()
sampleDepths=[]
for sample in sampleNames:
sampleDepths.append(str(sampleDepth[sample]))
MAPgenotypes=[ALPHABET[ i ] for i in MAPAssignment[0:totalSize] ]
#print MAPgenotypes
alt=determineAltBases( MAPgenotypes, [refsequence] )
numericalMAPgenotypes=[ numericalGenotypes(refsequence,alt,map_g) for map_g in MAPgenotypes ]
#print numericalMAPgenotypes
site_info="\t".join([pileup_column_chrom, str(pileup_column_pos+1), ".", refsequence,alt,qual,filter,infoString, "GT:GQ:DP"])
#zip the genotype assignment and its log-probability together
#genotypedata=zip(MAPgenotypes,sampleDepths,MAXvalues[0:totalSize])
#genotypedata=zip(MAPgenotypes,sampleDepths)
#genotypedata=zip(numericalMAPgenotypes,sampleDepths, MAXvalues_str[0:totalSize] )
genotypedata=zip(numericalMAPgenotypes,phred_str, sampleDepths, )
#genotypedata=zip(numericalMAPgenotypes,sampleDepths)
genotypedata=[ ":".join(list(i)) for i in genotypedata ]
#print genotypedata
#print genotypedata
#print sampleGenotypes
#print "\t".join( [pileup_column_chrom, str(pileup_column_pos), refsequence]) + "\t" +"\t".join(MAPgenotypes)
if int(pileup_column_pos) % 10000 == 0:
sys.stdout.write("processed 10 kbp ...\n")
#print "\t".join( [pileup_column_chrom, str(pileup_column_pos), refsequence]) + "\t" +"\t".join(numericalMAPgenotypes)
#print site_info
vcfgenotypeoutput="\t".join(genotypedata)
output=site_info+ "\t"+vcfgenotypeoutput
vcfh.write(output+"\n")
def main():
""" getting the git hash in python: http://stackoverflow.com/a/14989911/1735942"""
#label = subprocess.check_output(["git", "rev-parse", "HEAD"])
today=datetime.datetime.today()
datestr=today.strftime("20%y%m%d")
usage = "usage: %prog [options] my.bam "
parser = OptionParser(usage)
parser.add_option("--bed", type="string", dest="bedfile", help="bed file with coordinates")
parser.add_option("--tbf", type="string", dest="tbfile", help=" *.2bit file of the reference sequence")
parser.add_option("--ped", type="string", dest="pedfile", help= " pedfile of the samples you are analyzing")
parser.add_option("--min-nonref-count", type="int", dest="minAlt", default=2, help="minimum observation of nonref allele for genotype inference (default 2)")
parser.add_option("--mapq", dest="mapq", type="int", default=30, help="exclude read if mapping quality is less than Q default:30 ")
parser.add_option("--bq", dest="bq", type="int", default=25, help="exclude base if basequality is less than Q: default 25")
parser.add_option("--emitall", action="store_true", dest="emitall", help="emit all sites, even those without min alt threshold", default=False)
parser.add_option("--debug", action="store_true", dest="debug", default=False)
(options, args)=parser.parse_args()
commandline=";".join(sys.argv)
bamfile=args[0]
#print options.bedfile
bamfilebasename=return_file_basename(bamfile)
bedfilebasename=return_file_basename(options.bedfile)
vcfoutput=".".join(['pgmsnp',bamfilebasename,bedfilebasename, datestr, 'vcf'])
if options.debug == True:
prettybaseoutput=".".join(['pgmsnp',bamfilebasename, datestr,'pretty' ])
try:
os.remove(prettybaseoutput)
except OSError:
pass
prettyfh=open(prettybaseoutput, 'a')
vcfh=open(vcfoutput,'w')
DNA_ALPHABET= ['A', 'C', 'G' ,'T']
ALPHABET=['AA', 'AC', 'AG', 'AT', 'CC', 'CG', 'CT', 'GG', 'GT', 'TT']
twobit=bx.seq.twobit.TwoBitFile( open( options.tbfile ) )
bedobj=Bedfile(options.bedfile)
pybamfile=pysam.Samfile( bamfile, "rb" )
pedfile=Pedfile(options.pedfile)
pedfile.parsePedfile()
samplenames=pedfile.returnIndivids()
samplestring="\t".join(samplenames)
#vcf metainfolines
vcf_metalines=[]
vcf_metalines.append ( "##fileformat=VCFv4.1")
vcf_metalines.append( "##fileDate="+datestr )
vcf_metalines.append("##pgmsnp.py="+commandline)
#vcf_metalines.append("##version="+label.strip())
vcf_metalines.append("##reference="+options.tbfile)
vcf_metalines.append("##pedfile="+options.pedfile)
vcf_metalines.append("##bedfile="+options.bedfile)
vcf_metalines.append("##bamfile="+bamfile)
vcf_metalines.append("##INFO=<ID=NS,Number=1,Type=Integer,Description=\"Number of Samples With Data\">")
vcf_metalines.append( "##INFO=<ID=DP,Number=1,Type=Integer,Description=\"Total Depth\">" )
vcf_metalines.append( "##FORMAT=<ID=GT,Number=1,Type=String,Description=\"Genotype\">" )
vcf_metalines.append( "##FORMAT=<ID=GQ,Number=1,Type=Integer,Description=\"Genotype Quality\">" )
vcf_metalines.append( "##FORMAT=<ID=DP,Number=1,Type=Integer,Description=\"Read Depth\">" )
#vcf_metalines.apppend("##FILTER=<ID=LowQual,Description=\"Low quality\">")
vcf_column_headers=["#CHROM", "POS", "ID", "REF", "ALT", "QUAL", "FILTER", "INFO", "FORMAT",samplestring]
vcf_metalines.append( "\t".join(vcf_column_headers))
vcf_metaline_output="\n".join(vcf_metalines)
vcfh.write(vcf_metaline_output+"\n")
#print vcf_metaline_output
#print samplenames
#return the complete enumeration of all possible offspring genotype priors
punnetValues=returnPunnetValues(4)
# Pfactor gives us a pileup iterator
Pfactory=PileupFactory(pybamfile,bedobj)
##CHROM", "POS", "ID", "REF", "ALT", "QUAL", "FILTER", "INFO", "FORMAT",samplestring
for pileup_data_obj in Pfactory.yieldPileupData(options.mapq, options.bq):
refDepth=defaultdict(lambda: defaultdict(int))
altDepth=defaultdict(lambda: defaultdict(int))
pileup_column_pos=pileup_data_obj.getPileupColumnPos()
pileup_column_chrom=pileup_data_obj.getChrom()
refsequence=twobit[pileup_column_chrom][pileup_column_pos:pileup_column_pos+1] #this is the reference base
refDepth=defaultdict(lambda: defaultdict(int))
altDepth=defaultdict(lambda: defaultdict(int))
skipsite=False
#print 'Pileup position: ', pileup_data_obj, "\t".join( [pileup_column_chrom, str(pileup_column_pos), refsequence])
if refsequence not in DNA_ALPHABET: continue
#print
qual="."
filter='.'
siteDP="DP="+str(pileup_data_obj.getPileupDepth())
altDP=0
sampleDepth=defaultdict(int)
#lets make our genetic network here:
pgmNetwork=PgmNetworkFactory(options.pedfile,pileup_column_chrom,pileup_column_pos, refsequence,punnetValues)
totalSize=pgmNetwork.returnTotalSize()
#print "totalSize: ", totalSize
observedSamples=[]
sampleNames=set(pgmNetwork.getSampleNames())
#print "pgmNetwork factor list: "
#pgmNetwork.printFactorList()
pgmFactorList=pgmNetwork.getFactorList()
#print "++++"
for (sample, pileup_sample_data) in pileup_data_obj.yieldSamplePileupData():
sampleDepth[sample]=len(pileup_sample_data)
observedSamples.append(sample)
sample_idx=pgmNetwork.getSampleNamePedIndex(sample)
for data in pileup_sample_data:
if data.basecall != refsequence: altDepth[data.sample][data.basecall]+=1
if data.basecall == refsequence: refDepth[data.sample][data.basecall]+=1
value=calculateGLL(pileup_sample_data)
#pdb.set_trace()
pgmFactorList[sample_idx + totalSize].setVal(value)
#pdb.set_trace()
"""we initially get the log-likelihood, but go back to probablity space first """
#pgmFactorList[sample_idx + totalSize] = ExpFactor( pgmFactorList[sample_idx + totalSize] )
pgmFactorList[sample_idx + totalSize] = ExpFactorNormalize( pgmFactorList[sample_idx + totalSize] )
#print pgmFactorList[sample_idx + totalSize]
#GLFactor = Factor( [readVar, genoVar], [1,10], [], 'read_phenotype | genotype ')
#gPrior=LogFactor( returnGenotypePriorFounderFactor(sequence,['A','C','G','T'], genoVar) )
#print
""" in order for genotype inference to continue, minAlt value must be passed
If it isn't and emitall is True, then we do genotype inference. Otherwise continue"""
if sum(chain.from_iterable(d.itervalues() for d in altDepth.itervalues())) < options.minAlt and options.emitall == False:
continue
observedSamples=set(observedSamples)
unobservedSamples=sampleNames-observedSamples
unobservedIdxs=[ pgmNetwork.getSampleNamePedIndex(sample) for sample in unobservedSamples ]
#for samples lacking read data, delete them from the list of
for idx in unobservedIdxs:
#del pgmFactorList[idx + totalSize]
value=calculateNoObservationsGL()
#pdb.set_trace()
pgmFactorList[idx + totalSize].setVal(value)
#print pgmFactorList
#for f in pgmFactorList:
# print f
#continue
#print "====="
siteNS="NS="+str(len(observedSamples))
infoString=";".join([siteNS, siteDP])
#for f in pgmFactorList:
# print f
# print
#pdb.set_trace()
#cTree=CreatePrunedInitCtree(pgmFactorList)
#print 'cTree constructed, pruned, and initalized potential'
#G=nx.from_numpy_matrix( cTree.getEdges() )
#nx.draw_shell(G)
#plt.show()
#print cTree
#get the max marginal factors
if options.debug == True:
sys.stderr.write("computing exact marginals and joint from clique tree...\n")
(MAXMARGINALS,JOINT)=ComputeExactMarginalsBP(pgmFactorList, [], 1, 1)
sampleNames=pgmNetwork.returnGenotypeFactorIdxSampleNames()
#pdb.set_trace()
if options.debug == True:
posterior_genotypes_values(MAXMARGINALS, ALPHABET,samplenames, "\t".join([pileup_column_chrom, str(pileup_column_pos+1)]), prettyfh)
#this log normalizes the data
for m in MAXMARGINALS:
#print m
#print m.getVal()
#pdb.set_trace()
m.setVal( np.log( m.getVal() / np.sum(m.getVal() )) )
#m.setVal( np.log( lognormalize(m.getVal() ) ) )
#print np.sum( lognormalize(m.getVal() ) )
#print'lognormalize: ', lognormalize(m.getVal() )
#print m.getVal()
#print
# print m.getVal()
# print
#print "==="
#get the max value of each factor
#print len(MAXMARGINALS)
""" max values will hold the max; assuming its unique.
get a list of the the values as well """
#for m in MAXMARGINALS:
# print m
# print "total size: ", totalSize
# """ ask yourself, is the max unique? """
# print max(m.getVal().tolist())
# print
MAXvalues=[ max(m.getVal().tolist() ) for m in MAXMARGINALS ]
#pdb.set_trace()
#MAXvalues_phred = [ PhredScore(x) for x in MAXvalues ]
""" this code deals with calculating genotype quality GQ .... """
""" phred scale the error prob. so we take the 1-posterior, and phred scale it
Note we have expoentiate the values then take the complement, then phred scale """
phred_gq= [ round(PhredScore(x),2) for x in (1-np.exp(MAXvalues[0:totalSize])) ]
#MAXvalues_str= [str(x) for x in MAXvalues]
phred_str=[str(x) for x in phred_gq]
#print "maxvalues: ", MAXvalues[0:totalSize]
#this is the decoding, where we get teh assignment of the variable
# that has the max marginoal value
MAPAssignment=MaxDecoding( MAXMARGINALS )
#MAPAssignment=MaxDecodingNonUniq( MAXMARGINALS )
#pdb.set_trace()
#print "MAPAssignment: ", MAPAssignment[0:3]
#print totalSize
#for idx in range(totalSize):
# print ALPHABET[ MAPAssignment[idx] ]
#print MAPAssignment
#we convert from variable assignment in the factor to genotype space
sampleNames=pgmNetwork.returnGenotypeFactorIdxSampleNames()
sampleDepths=[]
for sample in sampleNames:
sampleDepths.append(str(sampleDepth[sample]))
MAPgenotypes=[ALPHABET[ i ] for i in MAPAssignment[0:totalSize] ]
#print MAPgenotypes
alt=determineAltBases( MAPgenotypes, [refsequence] )
dimension=np.prod(JOINT.getCard())
strides=variableStride(JOINT)
RR_genotype=refsequence+refsequence
idx=IndexToAssignment ( np.arange(np.prod(JOINT.getCard()) ), JOINT.getCard() )-1
g_idx= genotypeToIndex( RR_genotype, ploidy=2)
""" conviently enough, the index of the all ref assignment is genotype idx repeated however many times there are samples """
idx_string=str(g_idx)*totalSize
idx_numbr=int(idx_string)
allrefs_ass=generateAllReferenceGenotypesAssignment(g_idx, totalSize)
if np.array_equal(idx[idx_numbr], allrefs_ass) == False:
print "==="
print "value assignment doesn't equal all reference assignment"
print idx[idx_numbr]
print allrefs_ass
print "==="
sys.exit(1)
#idx_all_refs_ass=IndexOfAssignment(JOINT, strides, allref_ass )
else:
val=JOINT.getVal()[idx_numbr]
#pdb.set_trace()
""" Phred score takes in the error probability
In this case the error prob is the prob that everyone is homozygous reference
which is the val we get from the joint distribution factor above """
QUAL=round(PhredScore(val),2)
""" emit numerical genotypes as per vcf convention """
numericalMAPgenotypes=[ numericalGenotypes(refsequence,alt,map_g) for map_g in MAPgenotypes ]
#print numericalMAPgenotypes
site_info="\t".join([pileup_column_chrom, str(pileup_column_pos+1), ".",refsequence,alt,str(QUAL),filter,infoString, "GT:GQ:DP"])
#print site_info
#pdb.set_trace()
#site_info="\t".join([pileup_column_chrom, str(pileup_column_pos+1), ".", refsequence,alt,qual,filter,infoString, "GT:GQ:DP"])
#zip the genotype assignment and its log-probability together
#genotypedata=zip(MAPgenotypes,sampleDepths,MAXvalues[0:totalSize])
#genotypedata=zip(MAPgenotypes,sampleDepths)
#genotypedata=zip(numericalMAPgenotypes,sampleDepths, MAXvalues_str[0:totalSize] )
genotypedata=zip(numericalMAPgenotypes,phred_str, sampleDepths, )
#genotypedata=zip(numericalMAPgenotypes,sampleDepths)
genotypedata=[ ":".join(list(i)) for i in genotypedata ]
#print genotypedata
#print genotypedata
#print sampleGenotypes
#print "\t".join( [pileup_column_chrom, str(pileup_column_pos), refsequence]) + "\t" +"\t".join(MAPgenotypes)
if int(pileup_column_pos) % 10000 == 0:
sys.stdout.write("processed 10 kbp ...\n")
#print "\t".join( [pileup_column_chrom, str(pileup_column_pos), refsequence]) + "\t" +"\t".join(numericalMAPgenotypes)
#print site_info
vcfgenotypeoutput="\t".join(genotypedata)
output=site_info+ "\t"+vcfgenotypeoutput
vcfh.write(output+"\n")
