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"""Benchmarking module for AlignmentFile functionality"""
import os
import pysam
import unittest
from TestUtils import BAM_DATADIR, IS_PYTHON3, force_str, flatten_nested_list
import PileupTestUtils
class TestPileupReadSelection(unittest.TestCase):
'''test pileup functionality.'''
samfilename = os.path.join(BAM_DATADIR, "ex1.bam")
fastafilename = os.path.join(BAM_DATADIR, "ex1.fa")
def setUp(self):
self.samfile = pysam.AlignmentFile(self.samfilename)
self.fastafile = pysam.FastaFile(self.fastafilename)
def tearDown(self):
self.samfile.close()
self.fastafile.close()
def check_equal(self, references, iterator):
for x, column in enumerate(iterator):
v = references[x][:-1].split("\t")
self.assertEqual(
len(v), 6,
"expected 6 values, got {}".format(v))
(contig, pos, reference_base,
read_bases, read_qualities, alignment_mapping_qualities) \
= v
self.assertEqual(int(pos) - 1, column.reference_pos)
def test_samtools_stepper(self):
refs = force_str(
pysam.samtools.mpileup(
"-f", self.fastafilename,
self.samfilename)).splitlines(True)
iterator = self.samfile.pileup(
stepper="samtools",
fastafile=self.fastafile)
self.check_equal(refs, iterator)
def test_all_stepper(self):
refs = force_str(
pysam.samtools.mpileup(
"-f", self.fastafilename,
"-A", "-B",
self.samfilename)).splitlines(True)
iterator = self.samfile.pileup(
stepper="all",
fastafile=self.fastafile)
self.check_equal(refs, iterator)
def test_ignore_overlaps(self):
refs = force_str(
pysam.samtools.mpileup(
"-f", self.fastafilename,
"-A", "-B", "-x",
self.samfilename)).splitlines(True)
iterator = self.samfile.pileup(
stepper="all",
fastafile=self.fastafile,
ignore_overlaps=False)
self.check_equal(refs, iterator)
def test_samtools_stepper_mapping_quality_threshold(self):
refs = force_str(
pysam.samtools.mpileup(
"-f", self.fastafilename,
"--min-MQ", "15",
self.samfilename)).splitlines(True)
iterator = self.samfile.pileup(
stepper="samtools",
fastafile=self.fastafile,
min_mapping_quality=15)
self.check_equal(refs, iterator)
def test_samtools_stepper_base_quality_threshold(self):
refs = force_str(
pysam.samtools.mpileup(
"-f", self.fastafilename,
"--min-BQ", "20",
self.samfilename)).splitlines(True)
iterator = self.samfile.pileup(
stepper="samtools",
fastafile=self.fastafile,
min_base_quality=20)
self.check_equal(refs, iterator)
def test_samtools_stepper_ignore_orphans(self):
refs = force_str(
pysam.samtools.mpileup(
"-f", self.fastafilename,
"--count-orphans",
self.samfilename)).splitlines(True)
iterator = self.samfile.pileup(
stepper="samtools",
fastafile=self.fastafile,
ignore_orphans=False)
self.check_equal(refs, iterator)
def test_samtools_stepper_redo_baq(self):
refs = force_str(
pysam.samtools.mpileup(
"-f", self.fastafilename,
"--redo-BAQ",
self.samfilename)).splitlines(True)
iterator = self.samfile.pileup(
stepper="samtools",
fastafile=self.fastafile,
redo_baq=True)
self.check_equal(refs, iterator)
class TestPileupReadSelectionFastafile(TestPileupReadSelection):
'''test pileup functionality - backwards compatibility'''
samfilename = os.path.join(BAM_DATADIR, "ex1.bam")
fastafilename = os.path.join(BAM_DATADIR, "ex1.fa")
def setUp(self):
self.samfile = pysam.AlignmentFile(self.samfilename)
self.fastafile = pysam.Fastafile(self.fastafilename)
class TestPileupQueryPosition(unittest.TestCase):
filename = "test_query_position.bam"
def testPileup(self):
last = {}
with pysam.AlignmentFile(os.path.join(BAM_DATADIR, self.filename)) as inf:
for col in inf.pileup():
for r in col.pileups:
# print r.alignment.query_name
# print r.query_position, r.query_position_or_next, r.is_del
if r.is_del:
self.assertEqual(r.query_position, None)
self.assertEqual(r.query_position_or_next,
last[r.alignment.query_name] + 1)
else:
self.assertNotEqual(r.query_position, None)
last[r.alignment.query_name] = r.query_position
class TestPileupObjects(unittest.TestCase):
def setUp(self):
self.samfile = pysam.AlignmentFile(os.path.join(BAM_DATADIR, "ex1.bam"),
"rb")
def testPileupColumn(self):
for pcolumn1 in self.samfile.pileup(region="chr1:105-106"):
if pcolumn1.reference_pos == 104:
self.assertEqual(
pcolumn1.reference_id, 0,
"chromosome/target id mismatch in position 1: %s != %s" %
(pcolumn1.reference_id, 0))
self.assertEqual(
pcolumn1.reference_name, "chr1",
"chromosome mismatch in position 1: %s != %s" %
(pcolumn1.reference_name, "chr1"))
self.assertEqual(
pcolumn1.reference_pos, 105 - 1,
"position mismatch in position 1: %s != %s" %
(pcolumn1.reference_pos, 105 - 1))
self.assertEqual(
pcolumn1.nsegments, 1,
"# reads mismatch in position 1: %s != %s" %
(pcolumn1.nsegments, 1))
self.assertEqual(
len(pcolumn1.pileups), 1,
"# reads aligned to column mismatch in position 1"
": %s != %s" %
(len(pcolumn1.pileups), 1))
for pcolumn2 in self.samfile.pileup(region="chr2:1480-1481"):
if pcolumn2.reference_pos == 1479:
self.assertEqual(
pcolumn2.reference_id, 1,
"chromosome/target id mismatch in position 1: %s != %s" %
(pcolumn2.reference_id, 1))
self.assertEqual(
pcolumn2.reference_name, "chr2",
"chromosome mismatch in position 1: %s != %s" %
(pcolumn2.reference_name, "chr2"))
self.assertEqual(
pcolumn2.reference_pos, 1480 - 1,
"position mismatch in position 1: %s != %s" %
(pcolumn2.reference_pos, 1480 - 1))
self.assertEqual(
pcolumn2.nsegments, 12,
"# reads mismatch in position 1: %s != %s" %
(pcolumn2.nsegments, 12))
def tearDown(self):
self.samfile.close()
def testIteratorOutOfScope(self):
'''test if exception is raised if pileup col is accessed after
iterator is exhausted.'''
max_n = 0
for pileupcol in self.samfile.pileup():
if max_n < pileupcol.n:
max_col = pileupcol
max_n = pileupcol.n
self.assertRaises(ValueError, getattr, max_col, "pileups")
self.assertRaises(ValueError, max_col.get_query_sequences)
self.assertRaises(ValueError, max_col.get_num_aligned)
self.assertRaises(ValueError, max_col.get_query_qualities)
self.assertRaises(ValueError, max_col.get_mapping_qualities)
self.assertRaises(ValueError, max_col.get_query_positions)
self.assertRaises(ValueError, max_col.get_query_names)
class TestIteratorColumnBAM(unittest.TestCase):
'''test iterator column against contents of ex4.bam.'''
# note that samfile contains 1-based coordinates
# 1D means deletion with respect to reference sequence
#
mCoverages = {'chr1': [0] * 20 + [1] * 36 + [0] * (100 - 20 - 35),
'chr2': [0] * 20 + [1] * 35 + [0] * (100 - 20 - 35),
}
def setUp(self):
self.samfile = pysam.AlignmentFile(os.path.join(BAM_DATADIR, "ex4.bam"),
"rb")
def checkRange(self, contig, start=None, end=None, truncate=False):
'''compare results from iterator with those from samtools.'''
# check if the same reads are returned and in the same order
for column in self.samfile.pileup(
contig, start, end, truncate=truncate, min_base_quality=0):
if truncate:
self.assertGreaterEqual(column.reference_pos, start)
self.assertLess(column.reference_pos, end)
thiscov = len(column.pileups)
refcov = self.mCoverages[
self.samfile.getrname(column.reference_id)][column.reference_pos]
self.assertEqual(thiscov, refcov,
"wrong coverage at pos %s:%i %i should be %i" % (
self.samfile.getrname(column.reference_id),
column.reference_pos, thiscov, refcov))
def testIterateAll(self):
'''check random access per contig'''
self.checkRange(None)
def testIteratePerContig(self):
'''check random access per contig'''
for contig in self.samfile.references:
self.checkRange(contig)
def testIterateRanges(self):
'''check random access per range'''
for contig, length in zip(
self.samfile.references, self.samfile.lengths):
for start in range(1, length, 90):
# this includes empty ranges
self.checkRange(contig, start, start + 90)
def testInverse(self):
'''test the inverse, is point-wise pileup accurate.'''
for contig, refseq in list(self.mCoverages.items()):
refcolumns = sum(refseq)
for pos, refcov in enumerate(refseq):
columns = list(self.samfile.pileup(contig, pos, pos + 1))
if refcov == 0:
# if no read, no coverage
self.assertEqual(
len(columns),
refcov,
"wrong number of pileup columns returned for position %s:%i, %i should be %i" % (
contig, pos,
len(columns), refcov))
elif refcov == 1:
# one read, all columns of the read are returned
self.assertEqual(
len(columns),
refcolumns,
"pileup incomplete at position %i: got %i, expected %i " %
(pos, len(columns), refcolumns))
def testIterateTruncate(self):
'''check random access per range'''
for contig, length in zip(self.samfile.references,
self.samfile.lengths):
for start in range(1, length, 90):
# this includes empty ranges
self.checkRange(contig, start, start + 90, truncate=True)
def tearDown(self):
self.samfile.close()
class TestIteratorColumn2(unittest.TestCase):
'''test iterator column against contents of ex1.bam.'''
def setUp(self):
self.samfile = pysam.AlignmentFile(
os.path.join(BAM_DATADIR, "ex1.bam"),
"rb")
def testStart(self):
# print self.samfile.fetch().next().reference_start
# print self.samfile.pileup().next().reference_start
pass
def testTruncate(self):
'''see issue 107.'''
# note that ranges in regions start from 1
p = self.samfile.pileup(region='chr1:170-172', truncate=True)
columns = [x.reference_pos for x in p]
self.assertEqual(len(columns), 3)
self.assertEqual(columns, [169, 170, 171])
p = self.samfile.pileup('chr1', 169, 172, truncate=True)
columns = [x.reference_pos for x in p]
self.assertEqual(len(columns), 3)
self.assertEqual(columns, [169, 170, 171])
def testAccessOnClosedIterator(self):
'''see issue 131
Accessing pileup data after iterator has closed.
'''
pcolumn = self.samfile.pileup('chr1', 170, 180).__next__()
self.assertRaises(ValueError, getattr, pcolumn, "pileups")
def testStr(self):
'''test if PileupRead can be printed.'''
iter = self.samfile.pileup('chr1', 170, 180)
pcolumn = iter.__next__()
s = str(pcolumn)
self.assertEqual(len(s.split("\n")), 2)
@unittest.skipIf(not IS_PYTHON3,
"tests requires at least python3 for subprocess context manager")
class PileUpColumnTests(unittest.TestCase):
fn = os.path.join(BAM_DATADIR, "ex2.bam")
fn_fasta = os.path.join(BAM_DATADIR, "ex1.fa")
def test_pileup_depths_are_equal(self):
samtools_result = PileupTestUtils.build_depth_with_samtoolspipe(self.fn)
pysam_result = PileupTestUtils.build_depth_with_filter_with_pysam(self.fn)
self.assertEqual(pysam_result, samtools_result)
def test_pileup_query_bases_without_reference_are_equal(self):
samtools_result = PileupTestUtils.build_query_bases_with_samtoolspipe(self.fn)
pysam_result = PileupTestUtils.build_query_bases_with_pysam(self.fn)
self.assertEqual(["".join(x) for x in pysam_result], samtools_result)
def test_pileup_query_bases_with_reference_are_equal(self):
samtools_result = PileupTestUtils.build_query_bases_with_samtoolspipe(self.fn, "-f", self.fn_fasta)
with pysam.FastaFile(self.fn_fasta) as fasta:
pysam_result = PileupTestUtils.build_query_bases_with_pysam(self.fn, fastafile=fasta, stepper="samtools")
self.assertEqual(["".join(x) for x in pysam_result], samtools_result)
def test_pileup_query_qualities_are_equal(self):
samtools_result = PileupTestUtils.build_query_qualities_with_samtoolspipe(self.fn)
pysam_result = PileupTestUtils.build_query_qualities_with_pysam(self.fn)
pysam_result = [
[chr(min(126, x + 33)) for x in l] for l in pysam_result]
self.assertEqual("".join(flatten_nested_list(pysam_result)),
"".join(flatten_nested_list(samtools_result)))
def test_pileup_mapping_qualities_are_equal(self):
samtools_result = PileupTestUtils.build_mapping_qualities_with_samtoolspipe(self.fn)
pysam_result = PileupTestUtils.build_mapping_qualities_with_pysam(self.fn)
# convert to chars
pysam_result = [
[chr(min(126, x + 33)) for x in l] for l in pysam_result]
self.assertEqual("".join(flatten_nested_list(pysam_result)),
"".join(flatten_nested_list(samtools_result)))
def test_pileup_query_qualities_from_pileups_are_equal(self):
samtools_result = PileupTestUtils.build_query_qualities_with_samtoolspipe(self.fn)
pysam_result = PileupTestUtils.build_query_qualities_with_pysam_pileups(self.fn)
pysam_result = [
"".join([chr(min(126, x + 33)) for x in l]) for l in pysam_result]
self.assertEqual(pysam_result, samtools_result)
if __name__ == "__main__":
unittest.main()
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