def test1(self):
regions = []
add = regions.append
add(makeReadRegion(1, '2I', -1, -2))## an inserted I_1 before any M_1
add(makeReadRegion(0, '4M', 0, 3))
add(makeReadRegion(1, '2I', -1, -1)) ## an inserted I_1 after an M_1
add(makeReadRegion(1, '3I', -1, -1)) ## an inserted I_1 after an inserted I_1
## a gap
add(makeReadRegion(1, '1M', 5, 5)) ## a match I_1 after is an inserted I_1
add(makeReadRegion(1, '4I', 6, 5)) ## an inserted I_1 after a match I_1
## a gap here
add(makeReadRegion(0, '2M', 7, 8))
add(makeReadRegion(0, '2I', 9, 8)) ## an M_1 that is an insertion
add(makeReadRegion(1, '3I', 9, 8))
add(makeReadRegion(0, '4I', 9, 8)) ## an M_1 that is an insertion
add(makeReadRegion(1, '5I', 9, 8))
## a gap here
add(makeReadRegion(0, '5M', 10, 14))
cb = cigarbuilder.CigarBuilder()
cigar = cb.build(regions)
print(cigar)
self.assertEqual(cigar, [(1, 2), (0, 4), (1, 2), (1, 3), (2, 1),
(0, 1), (1, 4), (2, 1), (0, 2), (1, 2),
(1, 3), (1, 4), (1, 5), (2, 1), (0, 5)])
self.assertEqual(cigarutils.toString(cigar), '2I,4M,2I,3I,1D,1M,4I,1D,2M,2I,3I,4I,5I,1D,5M')
def logRegion(reg):
log(" op: %d.\n" % reg[0])
log(" region start: %d, end: %d, mpos: %d.\n"
% (reg[2], reg[3], reg[4]))
log(" region cigar: '%s'.\n" % cu.toString(reg[1]))
if len(reg) > 5:
log(" annotation: s: %d, i: %d, d: %d.\n" % (reg[5], reg[6], reg[7]))
def getReadOffset(rseq, pos):
'''
Given the position in the alignment coordinate, return its offset in read
'''
readPos = rseq.pos
#readLen = rseq.qlen
#qlen does not include soft-clipping, use rlen instead.
readLen = rseq.rlen
cigar = rseq.cigar
if (pos < readPos):
raise ValueError("position underflows")
offset = 0
curPos = readPos
for op, length in cigar:
if op == 0 or op == 7 or op == 8: # Match
if (pos < curPos + length): # (a) pos in this match
ret = offset + pos - curPos
if ret < readLen:
return ret
else:
msg = "cigar '%s' and length '%s' conflict in read '%s'"
cigarStr = cu.toString(cigar)
raise ValueError(msg % (cigarStr, readLen, rseq.qname))
else:
offset += length # (b) pos not in this match
curPos += length
elif op == 1 or op == 4: # Insertion and Soft Clipping
offset += length
elif op == 2 or op == 3: # Deletion or Splicing junction
curPos += length
else:
raise NotImplementedError("unknown op '%s' in read '%s'" %
(op, rseq.qname))
# In the case of offset == readLen, there may be an overflow
if offset > readLen:
msg = "cigar '%s' and length '%s' conflict in read '%s'"
cigarStr = cu.toString(cigar)
raise ValueError(msg % (cigarStr, readLen, rseq.qname))
if curPos > pos:
raise ValueError('position in deletion or splicing junction')
raise ValueError('position overflows')
def checkRegion(region):
'''sanity check for a region'''
assert len(region) >= 4
cigar = region[1]
nRefBases = 0
for op, length in cigar:
if op == 0 or op == 2 or op == 3:
nRefBases += length
if region[2]+ nRefBases - 1 != region[3]:
raise ValueError("Error: cigar '%s' conflicts with read region %d-%d."
% (cigarutils.toString(cigar), region[2], region[3]))
def checkRegion(region):
'''sanity check for a region'''
assert len(region) >= 4
cigar = region[1]
nRefBases = 0
for op, length in cigar:
if op == 0 or op == 2 or op == 3:
nRefBases += length
if region[2] + nRefBases - 1 != region[3]:
raise ValueError(
"Error: cigar '%s' conflicts with read region %d-%d." %
(cigarutils.toString(cigar), region[2], region[3]))
def test1(self):
regions = []
add = regions.append
## an inserted I_1 before any M_1
add(makeReadRegion(1, '2I', -1, -2))
add(makeReadRegion(0, '4M', 0, 3))
## an inserted I_1 after an M_1
add(makeReadRegion(1, '2I', -1, -1))
## an inserted I_1 after an inserted I_1
add(makeReadRegion(1, '3I', -1, -1))
## a gap
##
add(makeReadRegion(1, '1M', 5, 5))
## an inserted I_1 after a match I_1
add(makeReadRegion(1, '4I', 6, 5))
## a gap here
add(makeReadRegion(0, '2M', 7, 8))
## an M_1 that is an insertion
add(makeReadRegion(0, '2I', 9, 8))
add(makeReadRegion(1, '3I', 9, 8))
## an M_1 that is an insertion
add(makeReadRegion(0, '4I', 9, 8))
add(makeReadRegion(1, '5I', 9, 8))
## a gap here
add(makeReadRegion(0, '5M', 10, 14))
cb = cigarbuilder.CigarBuilder()
cigar = cb.build(regions)
print(cigar)
self.assertEqual(cigar, [(1, 2), (0, 4), (1, 2), (1, 3), (2, 1),
(1, 5), (2, 1), (0, 2), (1, 2), (1, 3),
(1, 4), (1, 5), (2, 1), (0, 5)])
self.assertEqual(cigarutils.toString(cigar),
'2I,4M,2I,3I,1D,1M,4I,1D,2M,2I,3I,4I,5I,1D,5M')
@author: Shunping Huang
'''
import unittest
import StringIO
import tempfile
import os
import pysam
from lapels import annotator as annot
from lapels import cigarutils
from modtools import mod
polish = lambda x: cigarutils.toString(cigarutils.simplify(x))
class Read:
'''Class for simulating reads from a bam file'''
def __init__(self, start, end, cigar=None, qlen=None):
self.qname = 'unknown'
self.pos = start
self.aend = end #one base after the actual end
self.tags = dict()
if cigar is None:
self.cigar = [(0, self.aend - self.pos)]
else:
self.cigar = cigar
if qlen is None:
Created on Oct 3, 2012
@author: Shunping Huang
'''
import unittest
import StringIO
import tempfile
import os
import pysam
from lapels import annotator as annot
from lapels import cigarutils
from modtools import mod
polish = lambda x: cigarutils.toString(cigarutils.simplify(x))
class Read:
'''Class for simulating reads from a bam file'''
def __init__(self, start, end, cigar=None, qlen=None):
self.qname = 'unknown'
self.pos = start
self.aend = end #one base after the actual end
self.tags = dict()
if cigar is None:
self.cigar = [(0, self.aend - self.pos)]
else:
self.cigar = cigar
if qlen is None:
self.qlen = 0
def testMakeCigar(self):
self.assertEqual(make(toString(self.cigar1)), self.cigar1)
self.assertEqual(make(toString(self.cigar2)), self.cigar2)
def testToString(self):
self.assertEqual(toString(self.cigar1),'15M,5I,10D,5M')
self.assertEqual(toString(self.cigar2),'10I,15M,5I,10D,5M,10I')
def execute(self):
'''The driver method for the module'''
self.logger.info("[%s]: %d read(s) found in BAM", self.chrom,
self.nReads)
if self.nReads == 0:
return 0
self.data = self.mod.data
self.modKeys = [tup[2] for tup in self.data]
self.posmap = self.mod.get_posmap(self.chrom)
count = 0
count2 = 0
if TESTING:
results = []
if not TESTING:
self.logger.info("[%s]: %3d%%", self.chrom,
count * 100 / self.nReads)
for rseq in self.inBam: # Annotate each reads
if rseq.is_unmapped: # write unmapped reads without changing
count += 1
if VERBOSITY > 1:
log("unmapped read name: %s\n" % rseq.qname)
if self.outBam is not None:
self.outBam.write(rseq)
continue
if VERBOSITY > 1:
log("read name: %s\n" % rseq.qname)
log("t. alignment pos: %d\n" % rseq.pos)
log("t. alignment cigar: '%s'\n\n" % cu.toString(rseq.cigar))
rseq.cigar = cu.simplify(rseq.cigar) # Simplify the cigar first.
regions = []
tregs = getTargetRegions(rseq)
for idx, treg in enumerate(tregs):
if treg[0] == 0 or treg[0] == 7 or treg[0] == 8: # Match
if VERBOSITY > 1:
log("process match\n")
log("t. region cigar(%d): %s\n"
% (idx, cu.toString([rseq.cigar[idx]])))
rreg = self.parseTargetRegion(treg, rseq)
if VERBOSITY > 1:
logRegion(rreg)
elif treg[0] == 2 or treg[0] == 3: # Deletion/Splice junction
rreg = (treg[0], )
else:
rreg = (1, [rseq.cigar[idx]], 0, -1, -1) # Insertion
regions.append(rreg)
if VERBOSITY > 1:
log("\nafter parsing match regions:\n")
log('\n'.join(map(str, regions)))
log("\n\n")
nRegions = len(regions)
assert nRegions == len(rseq.cigar)
for idx in range(nRegions):
op = regions[idx][0]
if op == 2 or op == 3: # Handle deletions and splicing
if VERBOSITY > 1:
log("process deletion/splicing junction\n")
log("t. region cigar(%d): %s\n"
% (idx, cu.toString([rseq.cigar[idx]])))
if ((idx > 0) and idx < (nRegions - 1) and
(regions[idx - 1][0] == 0) and
(regions[idx + 1][0] == 0)):
delta = regions[idx + 1][2] - regions[idx - 1][3] - 1
assert delta >= 0
if delta == 0:
rreg = (op, [],
regions[idx - 1][3] + 1,
regions[idx + 1][2] - 1,
-1)
else:
rreg = (op, [(op, delta)],
regions[idx - 1][3] + 1,
regions[idx + 1][2] - 1,
-1)
if VERBOSITY > 1:
logRegion(rreg)
else:
rreg = self.parseTargetRegion(tregs[idx], rseq)
if VERBOSITY > 1:
logRegion(rreg)
rreg = regionutils.modifyRegion(rreg)
if VERBOSITY > 1:
log("after modification\n")
logRegion(rreg)
regions[idx] = rreg
if VERBOSITY > 1:
log("\nafter parsing deletions/splicing junction regions:\n")
log('\n'.join(map(str, regions)))
log('\n\n')
cb = cigarbuilder.CigarBuilder()
# cigar=cb.build(regions)
for reg in regions:
cb.append(reg)
cigar = cb.cigar
# Fix the MIDM pattern
if re.match('.*\d*I,\d*D', cu.toString(cigar)) is not None:
if VERBOSITY > 1:
log("fix MIDM pattern: %s\n" % cu.toString(cigar))
# print("%d,%d,%d" %(nSNPs, nInsertions, nDeletions))
# print(regions)
for i in range(nRegions):
if tregs[i][0] == 1:
length = rseq.cigar[i][1]
assert length > 0
offset = getReadOffset(rseq, tregs[i][3]) + 1
ins = (rseq.seq[offset:offset + length])
if VERBOSITY > 1:
log('seq in insertion: %s\n' % ins)
if i > 0:
assert regions[i - 1][3] >= 0
loKey = regions[i - 1][3]
elif i < nRegions - 1 and regions[i + 1][2] >= length:
loKey = regions[i + 1][2] - length
else:
loKey = -1
if i < nRegions - 1:
assert regions[i + 1][2] >= 0
hiKey = regions[i + 1][2]
elif i > 0 and regions[i - 1][3] >= 0:
hiKey = regions[i - 1][3] + length
else:
hiKey = -1
# loKey = regions[i-1][3]
# hiKey = regions[i+1][2]
lo = bisect.bisect_left(self.modKeys, loKey)
hi = bisect.bisect_right(self.modKeys, hiKey)
if VERBOSITY > 1:
log('variants from %d-%d\n' % (loKey, hiKey))
for j in range(lo, hi):
log('%s\n' % str(self.data[j]))
isMatched = False
matchStart = -1
pivot = 0
isFailed = False
for j in range(lo, hi):
if self.data[j][0] != 'd':
# To activate MIDM pattern, the first instruction
# after insertion in read should be D.
isFailed = True
break
#continue
if matchStart == -1:
matchStart = int(self.data[j][2])
if ins[pivot] == self.data[j][3]:
pivot += 1
else:
break
if j == hi - 1 or pivot >= length:
isMatched = True
break
if isFailed:
continue
if isMatched:
if VERBOSITY > 1:
log('insertion matches gap from left\n')
log("before:\n")
logRegion(regions[i])
if pivot < length:
regions[i] = (0, [(0, pivot),
(1, length - pivot)],
matchStart,
matchStart + pivot - 1,
matchStart)
else:
regions[i] = (0, [(0, pivot)], matchStart,
matchStart + pivot - 1,
matchStart)
if VERBOSITY > 1:
log("after:\n")
logRegion(regions[i])
log("\n")
else:
pivot = length - 1
for j in range(hi - 1, lo - 1, -1):
if self.data[j][0] != 'd':
continue
if ins[pivot] == self.data[j][3]:
matchStart = int(self.data[j][2])
pivot -= 1
else:
break
if j == lo or pivot <= 0:
isMatched = True
break
if isMatched:
if VERBOSITY > 1:
log('insertion matches gap from right\n')
log("before:\n")
logRegion(regions[i])
if pivot >= 0:
regions[i] = (0, [(1, pivot + 1),
(0, length - 1 - pivot)],
matchStart,
matchStart + length - pivot - 2,
matchStart)
else:
regions[i] = (0, [(0, length - 1 - pivot)],
matchStart,
matchStart + length - pivot - 2,
matchStart)
if VERBOSITY > 1:
log("after:\n")
logRegion(regions[i])
log("\n")
else:
if VERBOSITY > 1:
log('insertion not matches\n')
cb = cigarbuilder.CigarBuilder()
# cigar=cb.build(regions)
for reg in regions:
cb.append(reg)
cigar = cb.cigar
# print(cu.toString(cigar))
if VERBOSITY > 1:
log("\nafter fixing special pattern:\n")
log('\n'.join(map(str, regions)))
log('\n\n')
nSNPs = 0
nInsertions = 0
nDeletions = 0
for reg in regions:
if len(reg) > 5:
nSNPs += reg[5]
nInsertions += reg[6]
nDeletions += reg[7]
## Set tags
tags = dict(rseq.tags)
self.setTag(tags, 's0', nSNPs)
self.setTag(tags, 'i0', nInsertions)
self.setTag(tags, 'd0', nDeletions)
self.setTag(tags, 'OC',
cu.toString(rseq.cigar).replace(',', ''))
self.setTag(tags, 'OM', tags['NM'])
del tags['NM'] # Delete the old 'NM' tag.
if nSNPs != 0 or nInsertions != 0 or nDeletions != 0:
count2 += 1
rseq.tags = [(key, tags[key]) for key in sorted(tags.keys())]
## Set pos to be the first M
pos = -1
for reg in regions:
if reg[4] >= 0:
pos = reg[4]
break
rseq.pos = pos
## Set cigar
rseq.cigar = cu.simplify(cigar)
if self.outBam is not None:
self.outBam.write(rseq)
if TESTING:
results.append((rseq.cigar, rseq.pos, nSNPs, nInsertions,
nDeletions))
if VERBOSITY > 1:
log("output read pos: %d\n" % rseq.pos)
log("output read cigar: '%s'\n"
% cu.toString(rseq.cigar))
log("output annotation: s: %d, i: %d, d: %d.\n"
"========================\n\n" %
(nSNPs, nInsertions, nDeletions))
count += 1
if not TESTING and count % 100000 == 0:
self.logger.info("[%s]: %3d%%", self.chrom,
count * 100 / self.nReads)
if not TESTING:
self.logger.info("[%s]: %3d%%", self.chrom,
count * 100 / self.nReads)
self.logger.info("[%s]: %d read(s) written to file", self.chrom,
count)
self.logger.info("[%s]: %d read(s) have variants", self.chrom,
count2)
else:
return results
return count
