def _varscan_work(align_bams, ref_file, config, target_regions, out_file):
"""Perform SNP and indel genotyping with VarScan.
"""
max_read_depth = "1000"
version = programs.jar_versioner("varscan", "VarScan")(config)
if version < "v2.3.5":
raise IOError("Please install version 2.3.5 or better of VarScan with support "
"for multisample calling and indels in VCF format.")
varscan_jar = config_utils.get_jar("VarScan",
config_utils.get_program("varscan", config, "dir"))
resources = config_utils.get_resources("varscan", config)
jvm_opts = " ".join(resources.get("jvm_opts", ["-Xmx750m", "-Xmx2g"]))
sample_list = _create_sample_list(align_bams, out_file)
mpileup = samtools.prep_mpileup(align_bams, ref_file, max_read_depth, config,
target_regions=target_regions, want_bcf=False)
cmd = ("{mpileup} "
"| java {jvm_opts} -jar {varscan_jar} mpileup2cns --min-coverage 5 --p-value 0.98 "
" --vcf-sample-list {sample_list} --output-vcf --variants "
"> {out_file}")
cmd = cmd.format(**locals())
do.run(cmd, "Varscan".format(**locals()), None,
[do.file_exists(out_file)])
os.remove(sample_list)
# VarScan can create completely empty files in regions without
# variants, so we create a correctly formatted empty file
if os.path.getsize(out_file) == 0:
write_empty_vcf(out_file)
def _varscan_work(align_bams, ref_file, items, target_regions, out_file):
"""Perform SNP and indel genotyping with VarScan.
"""
config = items[0]["config"]
max_read_depth = "1000"
version = programs.jar_versioner("varscan", "VarScan")(config)
if version < "v2.3.5":
raise IOError("Please install version 2.3.5 or better of VarScan with support "
"for multisample calling and indels in VCF format.")
varscan_jar = config_utils.get_jar("VarScan",
config_utils.get_program("varscan", config, "dir"))
jvm_opts = _get_varscan_opts(config)
sample_list = _create_sample_list(align_bams, out_file)
mpileup = samtools.prep_mpileup(align_bams, ref_file, max_read_depth, config,
target_regions=target_regions, want_bcf=False)
# VarScan fails to generate a header on files that start with
# zerocoverage calls; strip these with grep, we're not going to
# call on them
remove_zerocoverage = "grep -v -P '\t0\t\t$'"
cmd = ("{mpileup} | {remove_zerocoverage} "
"| java {jvm_opts} -jar {varscan_jar} mpileup2cns --min-coverage 5 --p-value 0.98 "
" --vcf-sample-list {sample_list} --output-vcf --variants "
"> {out_file}")
cmd = cmd.format(**locals())
do.run(cmd, "Varscan".format(**locals()), None,
[do.file_exists(out_file)])
os.remove(sample_list)
# VarScan can create completely empty files in regions without
# variants, so we create a correctly formatted empty file
if os.path.getsize(out_file) == 0:
write_empty_vcf(out_file)
def _varscan_work(align_bams, ref_file, config, target_regions, out_file):
"""Perform SNP and indel genotyping with VarScan.
"""
max_read_depth = "1000"
version = programs.jar_versioner("varscan", "VarScan")(config)
if version < "v2.3.5":
raise IOError(
"Please install version 2.3.5 or better of VarScan with support "
"for multisample calling and indels in VCF format.")
varscan_jar = config_utils.get_jar(
"VarScan", config_utils.get_program("varscan", config, "dir"))
resources = config_utils.get_resources("varscan", config)
jvm_opts = " ".join(resources.get("jvm_opts", ["-Xmx750m", "-Xmx2g"]))
sample_list = _create_sample_list(align_bams, out_file)
mpileup = samtools.prep_mpileup(align_bams,
ref_file,
max_read_depth,
config,
target_regions=target_regions,
want_bcf=False)
cmd = (
"{mpileup} "
"| java {jvm_opts} -jar {varscan_jar} mpileup2cns --min-coverage 5 --p-value 0.98 "
" --vcf-sample-list {sample_list} --output-vcf --variants "
"> {out_file}")
cmd = cmd.format(**locals())
do.run(cmd, "Varscan".format(**locals()), None, [do.file_exists(out_file)])
os.remove(sample_list)
# VarScan can create completely empty files in regions without
# variants, so we create a correctly formatted empty file
if os.path.getsize(out_file) == 0:
write_empty_vcf(out_file)
def _varscan_work(align_bams, ref_file, items, target_regions, out_file):
"""Perform SNP and indel genotyping with VarScan.
"""
config = items[0]["config"]
orig_out_file = out_file
out_file = orig_out_file.replace(".vcf.gz", ".vcf")
max_read_depth = "1000"
version = programs.jar_versioner("varscan", "VarScan")(config)
if version < "v2.3.6":
raise IOError("Please install version 2.3.6 or better of VarScan"
" with support for multisample calling and indels"
" in VCF format.")
varscan_jar = config_utils.get_jar(
"VarScan", config_utils.get_program("varscan", config, "dir"))
sample_list = _create_sample_list(align_bams, out_file)
mpileup = samtools.prep_mpileup(align_bams,
ref_file,
config,
max_read_depth,
target_regions=target_regions,
want_bcf=False)
# VarScan fails to generate a header on files that start with
# zerocoverage calls; strip these with grep, we're not going to
# call on them
remove_zerocoverage = "grep -v -P '\t0\t\t$'"
# write a temporary mpileup file so we can check if empty
mpfile = "%s.mpileup" % os.path.splitext(out_file)[0]
with file_transaction(config, mpfile) as mpfile_tx:
cmd = ("{mpileup} | {remove_zerocoverage} > {mpfile_tx}")
do.run(cmd.format(**locals()), "mpileup for Varscan")
if os.path.getsize(mpfile) == 0:
write_empty_vcf(out_file)
else:
with tx_tmpdir(items[0]) as tmp_dir:
jvm_opts = _get_varscan_opts(config, tmp_dir)
fix_ambig = vcfutils.fix_ambiguous_cl()
cmd = (
"cat {mpfile} "
"| java {jvm_opts} -jar {varscan_jar} mpileup2cns --min-coverage 5 --p-value 0.98 "
" --vcf-sample-list {sample_list} --output-vcf --variants "
"| {fix_ambig} | vcfuniqalleles > {out_file}")
do.run(cmd.format(**locals()), "Varscan", None,
[do.file_exists(out_file)])
os.remove(sample_list)
os.remove(mpfile)
# VarScan can create completely empty files in regions without
# variants, so we create a correctly formatted empty file
if os.path.getsize(out_file) == 0:
write_empty_vcf(out_file)
else:
freebayes.clean_vcf_output(out_file, _clean_varscan_line, config)
if orig_out_file.endswith(".gz"):
vcfutils.bgzip_and_index(out_file, config)
def _varscan_work(align_bams, ref_file, items, target_regions, out_file):
"""Perform SNP and indel genotyping with VarScan.
"""
config = items[0]["config"]
orig_out_file = out_file
out_file = orig_out_file.replace(".vcf.gz", ".vcf")
max_read_depth = "1000"
version = programs.jar_versioner("varscan", "VarScan")(config)
if version < "v2.3.6":
raise IOError("Please install version 2.3.6 or better of VarScan"
" with support for multisample calling and indels"
" in VCF format.")
varscan_jar = config_utils.get_jar("VarScan",
config_utils.get_program("varscan", config, "dir"))
sample_list = _create_sample_list(align_bams, out_file)
mpileup = samtools.prep_mpileup(align_bams, ref_file, config, max_read_depth,
target_regions=target_regions, want_bcf=False)
# VarScan fails to generate a header on files that start with
# zerocoverage calls; strip these with grep, we're not going to
# call on them
remove_zerocoverage = "grep -v -P '\t0\t\t$'"
# write a temporary mpileup file so we can check if empty
mpfile = "%s.mpileup" % os.path.splitext(out_file)[0]
with file_transaction(config, mpfile) as mpfile_tx:
cmd = ("{mpileup} | {remove_zerocoverage} > {mpfile_tx}")
do.run(cmd.format(**locals()), "mpileup for Varscan")
if os.path.getsize(mpfile) == 0:
write_empty_vcf(out_file)
else:
with tx_tmpdir(items[0]) as tmp_dir:
jvm_opts = _get_varscan_opts(config, tmp_dir)
fix_ambig = vcfutils.fix_ambiguous_cl()
cmd = ("cat {mpfile} "
"| java {jvm_opts} -jar {varscan_jar} mpileup2cns --min-coverage 5 --p-value 0.98 "
" --vcf-sample-list {sample_list} --output-vcf --variants "
"| {fix_ambig} | vcfuniqalleles > {out_file}")
do.run(cmd.format(**locals()), "Varscan", None,
[do.file_exists(out_file)])
os.remove(sample_list)
os.remove(mpfile)
# VarScan can create completely empty files in regions without
# variants, so we create a correctly formatted empty file
if os.path.getsize(out_file) == 0:
write_empty_vcf(out_file)
else:
freebayes.clean_vcf_output(out_file, _clean_varscan_line, config)
if orig_out_file.endswith(".gz"):
vcfutils.bgzip_and_index(out_file, config)
def _varscan_work(align_bams, ref_file, items, target_regions, out_file):
"""Perform SNP and indel genotyping with VarScan.
"""
config = items[0]["config"]
orig_out_file = out_file
out_file = orig_out_file.replace(".vcf.gz", ".vcf")
max_read_depth = "1000"
sample_list = _create_sample_list(align_bams, out_file)
mpileup = samtools.prep_mpileup(align_bams,
ref_file,
config,
max_read_depth,
target_regions=target_regions,
want_bcf=False)
# VarScan fails to generate a header on files that start with
# zerocoverage calls; strip these with grep, we're not going to
# call on them
remove_zerocoverage = r"{ ifne grep -v -P '\t0\t\t$' || true; }"
# we use ifne from moreutils to ensure we process only on files with input, skipping otherwise
# http://manpages.ubuntu.com/manpages/natty/man1/ifne.1.html
with tx_tmpdir(items[0]) as tmp_dir:
jvm_opts = _get_jvm_opts(config, tmp_dir)
opts = " ".join(_varscan_options_from_config(config))
min_af = float(
utils.get_in(config,
("algorithm", "min_allele_fraction"), 10)) / 100.0
fix_ambig_ref = vcfutils.fix_ambiguous_cl()
fix_ambig_alt = vcfutils.fix_ambiguous_cl(5)
py_cl = os.path.join(os.path.dirname(sys.executable), "py")
export = utils.local_path_export()
cmd = (
"{export} {mpileup} | {remove_zerocoverage} | "
"ifne varscan {jvm_opts} mpileup2cns {opts} "
"--vcf-sample-list {sample_list} --min-var-freq {min_af} --output-vcf --variants | "
"""{py_cl} -x 'bcbio.variation.vcfutils.add_contig_to_header(x, "{ref_file}")' | """
"{py_cl} -x 'bcbio.variation.varscan.fix_varscan_output(x)' | "
"{fix_ambig_ref} | {fix_ambig_alt} | ifne vcfuniqalleles > {out_file}"
)
do.run(cmd.format(**locals()), "Varscan", None,
[do.file_exists(out_file)])
os.remove(sample_list)
# VarScan can create completely empty files in regions without
# variants, so we create a correctly formatted empty file
if os.path.getsize(out_file) == 0:
write_empty_vcf(out_file)
if orig_out_file.endswith(".gz"):
vcfutils.bgzip_and_index(out_file, config)
def _varscan_work(align_bams, ref_file, items, target_regions, out_file):
"""Perform SNP and indel genotyping with VarScan.
"""
config = items[0]["config"]
orig_out_file = out_file
out_file = orig_out_file.replace(".vcf.gz", ".vcf")
max_read_depth = "1000"
sample_list = _create_sample_list(align_bams, out_file)
mpileup = samtools.prep_mpileup(align_bams, ref_file, config, max_read_depth,
target_regions=target_regions, want_bcf=False)
# VarScan fails to generate a header on files that start with
# zerocoverage calls; strip these with grep, we're not going to
# call on them
remove_zerocoverage = r"{ ifne grep -v -P '\t0\t\t$' || true; }"
# we use ifne from moreutils to ensure we process only on files with input, skipping otherwise
# http://manpages.ubuntu.com/manpages/natty/man1/ifne.1.html
with tx_tmpdir(items[0]) as tmp_dir:
jvm_opts = _get_jvm_opts(config, tmp_dir)
opts = " ".join(_varscan_options_from_config(config))
min_af = float(utils.get_in(config, ("algorithm", "min_allele_fraction"), 10)) / 100.0
fix_ambig_ref = vcfutils.fix_ambiguous_cl()
fix_ambig_alt = vcfutils.fix_ambiguous_cl(5)
py_cl = os.path.join(os.path.dirname(sys.executable), "py")
export = utils.local_path_export()
cmd = ("{export} {mpileup} | {remove_zerocoverage} | "
"ifne varscan {jvm_opts} mpileup2cns {opts} "
"--vcf-sample-list {sample_list} --min-var-freq {min_af} --output-vcf --variants | "
"""{py_cl} -x 'bcbio.variation.vcfutils.add_contig_to_header(x, "{ref_file}")' | """
"{py_cl} -x 'bcbio.variation.varscan.fix_varscan_output(x)' | "
"{fix_ambig_ref} | {fix_ambig_alt} | ifne vcfuniqalleles > {out_file}")
do.run(cmd.format(**locals()), "Varscan", None,
[do.file_exists(out_file)])
os.remove(sample_list)
# VarScan can create completely empty files in regions without
# variants, so we create a correctly formatted empty file
if os.path.getsize(out_file) == 0:
write_empty_vcf(out_file)
if orig_out_file.endswith(".gz"):
vcfutils.bgzip_and_index(out_file, config)
def _varscan_work(align_bams, ref_file, items, target_regions, out_file):
"""Perform SNP and indel genotyping with VarScan.
"""
config = items[0]["config"]
max_read_depth = "1000"
version = programs.jar_versioner("varscan", "VarScan")(config)
if version < "v2.3.6":
raise IOError("Please install version 2.3.6 or better of VarScan"
" with support for multisample calling and indels"
" in VCF format.")
varscan_jar = config_utils.get_jar(
"VarScan", config_utils.get_program("varscan", config, "dir"))
jvm_opts = _get_varscan_opts(config)
sample_list = _create_sample_list(align_bams, out_file)
mpileup = samtools.prep_mpileup(align_bams,
ref_file,
max_read_depth,
config,
target_regions=target_regions,
want_bcf=False)
# VarScan fails to generate a header on files that start with
# zerocoverage calls; strip these with grep, we're not going to
# call on them
remove_zerocoverage = "grep -v -P '\t0\t\t$'"
cmd = (
"{mpileup} | {remove_zerocoverage} "
"| java {jvm_opts} -jar {varscan_jar} mpileup2cns --min-coverage 5 --p-value 0.98 "
" --vcf-sample-list {sample_list} --output-vcf --variants "
"> {out_file}")
cmd = cmd.format(**locals())
do.run(cmd, "Varscan".format(**locals()), None, [do.file_exists(out_file)])
os.remove(sample_list)
# VarScan can create completely empty files in regions without
# variants, so we create a correctly formatted empty file
if os.path.getsize(out_file) == 0:
write_empty_vcf(out_file)
def _varscan_paired(align_bams, ref_file, items, target_regions, out_file):
"""Run a paired VarScan analysis, also known as "somatic". """
max_read_depth = "1000"
config = items[0]["config"]
version = programs.jar_versioner("varscan", "VarScan")(config)
if LooseVersion(version) < LooseVersion("v2.3.6"):
raise IOError(
"Please install version 2.3.6 or better of VarScan with support "
"for multisample calling and indels in VCF format.")
varscan_jar = config_utils.get_jar(
"VarScan",
config_utils.get_program("varscan", config, "dir"))
remove_zerocoverage = "grep -v -P '\t0\t\t$'"
# No need for names in VarScan, hence the "_"
paired = get_paired_bams(align_bams, items)
if not paired.normal_bam:
raise ValueError("Require both tumor and normal BAM files for VarScan cancer calling")
if not file_exists(out_file):
orig_out_file = out_file
out_file = orig_out_file.replace(".vcf.gz", ".vcf")
base, ext = utils.splitext_plus(out_file)
cleanup_files = []
for fname, mpext in [(paired.normal_bam, "normal"), (paired.tumor_bam, "tumor")]:
mpfile = "%s-%s.mpileup" % (base, mpext)
cleanup_files.append(mpfile)
with file_transaction(mpfile) as mpfile_tx:
mpileup = samtools.prep_mpileup([fname], ref_file,
max_read_depth, config,
target_regions=target_regions,
want_bcf=False)
cmd = "{mpileup} > {mpfile_tx}"
cmd = cmd.format(**locals())
do.run(cmd, "samtools mpileup".format(**locals()), None,
[do.file_exists(mpfile_tx)])
# Sometimes mpileup writes an empty file: in this case we
# just skip the rest of the analysis (VarScan will hang otherwise)
if any(os.stat(filename).st_size == 0 for filename in cleanup_files):
write_empty_vcf(orig_out_file, config)
return
# First index is normal, second is tumor
normal_tmp_mpileup = cleanup_files[0]
tumor_tmp_mpileup = cleanup_files[1]
indel_file = base + ".indel.vcf"
snp_file = base + ".snp.vcf"
cleanup_files.append(indel_file)
cleanup_files.append(snp_file)
with file_transaction(indel_file, snp_file) as (tx_indel, tx_snp):
with utils.curdir_tmpdir(items[0]) as tmp_dir:
jvm_opts = _get_varscan_opts(config, tmp_dir)
varscan_cmd = ("java {jvm_opts} -jar {varscan_jar} somatic"
" {normal_tmp_mpileup} {tumor_tmp_mpileup} {base}"
" --output-vcf --min-coverage 5 --p-value 0.98 "
"--strand-filter 1 ")
# add minimum AF
if "--min-var-freq" not in varscan_cmd:
min_af = float(utils.get_in(paired.tumor_config, ("algorithm",
"min_allele_fraction"),10)) / 100.0
varscan_cmd += "--min-var-freq {min_af} "
do.run(varscan_cmd.format(**locals()), "Varscan", None, None)
# VarScan files need to be corrected to match the VCF specification
# We do this before combining them otherwise merging may fail
# if there are invalid records
to_combine = []
if do.file_exists(snp_file):
to_combine.append(snp_file)
_fix_varscan_vcf(snp_file, paired.normal_name, paired.tumor_name)
if do.file_exists(indel_file):
to_combine.append(indel_file)
_fix_varscan_vcf(indel_file, paired.normal_name, paired.tumor_name)
if not to_combine:
write_empty_vcf(orig_out_file, config)
return
out_file = combine_variant_files([snp_file, indel_file],
out_file, ref_file, config,
region=target_regions)
# Remove cleanup files
for extra_file in cleanup_files:
for ext in ["", ".gz", ".gz.tbi"]:
if os.path.exists(extra_file + ext):
os.remove(extra_file + ext)
if os.path.getsize(out_file) == 0:
write_empty_vcf(out_file)
if orig_out_file.endswith(".gz"):
out_file = bgzip_and_index(out_file, config)
_add_reject_flag(out_file, config)
def _varscan_paired(align_bams, ref_file, items, target_regions, out_file):
"""Run a paired VarScan analysis, also known as "somatic". """
max_read_depth = "1000"
config = items[0]["config"]
version = programs.jar_versioner("varscan", "VarScan")(config)
if LooseVersion(version) < LooseVersion("v2.3.6"):
raise IOError(
"Please install version 2.3.6 or better of VarScan with support "
"for multisample calling and indels in VCF format.")
varscan_jar = config_utils.get_jar(
"VarScan", config_utils.get_program("varscan", config, "dir"))
remove_zerocoverage = "grep -v -P '\t0\t\t$'"
# No need for names in VarScan, hence the "_"
paired = get_paired_bams(align_bams, items)
if not paired.normal_bam:
raise ValueError(
"Require both tumor and normal BAM files for VarScan cancer calling"
)
if not file_exists(out_file):
base, ext = os.path.splitext(out_file)
cleanup_files = []
for fname, mpext in [(paired.normal_bam, "normal"),
(paired.tumor_bam, "tumor")]:
mpfile = "%s-%s.mpileup" % (base, mpext)
cleanup_files.append(mpfile)
with file_transaction(mpfile) as mpfile_tx:
mpileup = samtools.prep_mpileup([fname],
ref_file,
max_read_depth,
config,
target_regions=target_regions,
want_bcf=False)
cmd = "{mpileup} > {mpfile_tx}"
cmd = cmd.format(**locals())
do.run(cmd, "samtools mpileup".format(**locals()), None,
[do.file_exists(mpfile_tx)])
# Sometimes mpileup writes an empty file: in this case we
# just skip the rest of the analysis (VarScan will hang otherwise)
if any(os.stat(filename).st_size == 0 for filename in cleanup_files):
write_empty_vcf(out_file)
return
# First index is normal, second is tumor
normal_tmp_mpileup = cleanup_files[0]
tumor_tmp_mpileup = cleanup_files[1]
jvm_opts = _get_varscan_opts(config)
varscan_cmd = ("java {jvm_opts} -jar {varscan_jar} somatic"
" {normal_tmp_mpileup} {tumor_tmp_mpileup} {base}"
" --output-vcf --min-coverage 5 --p-value 0.98 "
"--strand-filter 1 ")
indel_file = base + ".indel.vcf"
snp_file = base + ".snp.vcf"
cleanup_files.append(indel_file)
cleanup_files.append(snp_file)
to_combine = []
with file_transaction(indel_file, snp_file) as (tx_indel, tx_snp):
varscan_cmd = varscan_cmd.format(**locals())
do.run(varscan_cmd, "Varscan".format(**locals()), None, None)
# VarScan files need to be corrected to match the VCF specification
# We do this before combining them otherwise merging may fail
# if there are invalid records
if do.file_exists(snp_file):
to_combine.append(snp_file)
_fix_varscan_vcf(snp_file, paired.normal_name, paired.tumor_name)
if do.file_exists(indel_file):
to_combine.append(indel_file)
_fix_varscan_vcf(indel_file, paired.normal_name, paired.tumor_name)
if not to_combine:
write_empty_vcf(out_file)
return
out_file = combine_variant_files([snp_file, indel_file],
out_file,
ref_file,
config,
region=target_regions)
# Remove cleanup files
for extra_file in cleanup_files:
os.remove(extra_file)
if os.path.getsize(out_file) == 0:
write_empty_vcf(out_file)
def _varscan_paired(align_bams, ref_file, items, target_regions, out_file):
"""Run a paired VarScan analysis, also known as "somatic". """
max_read_depth = "1000"
config = items[0]["config"]
version = programs.jar_versioner("varscan", "VarScan")(config)
if LooseVersion(version) < LooseVersion("v2.3.6"):
raise IOError(
"Please install version 2.3.6 or better of VarScan with support "
"for multisample calling and indels in VCF format.")
varscan_jar = config_utils.get_jar(
"VarScan",
config_utils.get_program("varscan", config, "dir"))
remove_zerocoverage = "grep -v -P '\t0\t\t$'"
# No need for names in VarScan, hence the "_"
tumor_bam, tumor_name, normal_bam, normal_name = get_paired_bams(
align_bams, items)
if not file_exists(out_file):
base, ext = os.path.splitext(out_file)
cleanup_files = []
for fname, mpext in [(normal_bam, "normal"), (tumor_bam, "tumor")]:
mpfile = "%s-%s.mpileup" % (base, mpext)
cleanup_files.append(mpfile)
with file_transaction(mpfile) as mpfile_tx:
mpileup = samtools.prep_mpileup([fname], ref_file,
max_read_depth, config,
target_regions=target_regions,
want_bcf=False)
cmd = "{mpileup} > {mpfile_tx}"
cmd = cmd.format(**locals())
do.run(cmd, "samtools mpileup".format(**locals()), None,
[do.file_exists(mpfile_tx)])
# Sometimes mpileup writes an empty file: in this case we
# just skip the rest of the analysis (VarScan will hang otherwise)
if any(os.stat(filename).st_size == 0 for filename in cleanup_files):
write_empty_vcf(out_file)
return
# First index is normal, second is tumor
normal_tmp_mpileup = cleanup_files[0]
tumor_tmp_mpileup = cleanup_files[1]
jvm_opts = _get_varscan_opts(config)
varscan_cmd = ("java {jvm_opts} -jar {varscan_jar} somatic"
" {normal_tmp_mpileup} {tumor_tmp_mpileup} {base}"
" --output-vcf --min-coverage 5 --p-value 0.98")
indel_file = base + ".indel.vcf"
snp_file = base + ".snp.vcf"
cleanup_files.append(indel_file)
cleanup_files.append(snp_file)
to_combine = []
with file_transaction(indel_file, snp_file) as (tx_indel, tx_snp):
varscan_cmd = varscan_cmd.format(**locals())
do.run(varscan_cmd, "Varscan".format(**locals()), None,
None)
# VarScan files need to be corrected to match the VCF specification
# We do this before combining them otherwise merging may fail
# if there are invalid records
if do.file_exists(snp_file):
to_combine.append(snp_file)
_fix_varscan_vcf(snp_file, normal_name, tumor_name)
if do.file_exists(indel_file):
to_combine.append(indel_file)
_fix_varscan_vcf(indel_file, normal_name, tumor_name)
if not to_combine:
write_empty_vcf(out_file)
return
out_file = combine_variant_files([snp_file, indel_file],
out_file, ref_file, config,
region=target_regions)
# Remove cleanup files
for extra_file in cleanup_files:
os.remove(extra_file)
if os.path.getsize(out_file) == 0:
write_empty_vcf(out_file)
def _varscan_paired(align_bams, ref_file, items, target_regions, out_file):
"""Run a paired VarScan analysis, also known as "somatic". """
max_read_depth = "1000"
config = items[0]["config"]
version = programs.jar_versioner("varscan", "VarScan")(config)
if LooseVersion(version) < LooseVersion("v2.3.6"):
raise IOError(
"Please install version 2.3.6 or better of VarScan with support "
"for multisample calling and indels in VCF format.")
varscan_jar = config_utils.get_jar(
"VarScan", config_utils.get_program("varscan", config, "dir"))
remove_zerocoverage = "grep -v -P '\t0\t\t$'"
# No need for names in VarScan, hence the "_"
paired = get_paired_bams(align_bams, items)
if not paired.normal_bam:
affected_batch = items[0]["metadata"]["batch"]
message = ("Batch {} requires both tumor and normal BAM files for"
" VarScan cancer calling").format(affected_batch)
raise ValueError(message)
if not file_exists(out_file):
orig_out_file = out_file
out_file = orig_out_file.replace(".vcf.gz", ".vcf")
base, ext = utils.splitext_plus(out_file)
cleanup_files = []
for fname, mpext in [(paired.normal_bam, "normal"),
(paired.tumor_bam, "tumor")]:
mpfile = "%s-%s.mpileup" % (base, mpext)
cleanup_files.append(mpfile)
with file_transaction(config, mpfile) as mpfile_tx:
mpileup = samtools.prep_mpileup([fname],
ref_file,
config,
max_read_depth,
target_regions=target_regions,
want_bcf=False)
cmd = "{mpileup} > {mpfile_tx}"
cmd = cmd.format(**locals())
do.run(cmd, "samtools mpileup".format(**locals()), None,
[do.file_exists(mpfile_tx)])
# Sometimes mpileup writes an empty file: in this case we
# just skip the rest of the analysis (VarScan will hang otherwise)
if any(os.stat(filename).st_size == 0 for filename in cleanup_files):
write_empty_vcf(orig_out_file, config)
return
# First index is normal, second is tumor
normal_tmp_mpileup = cleanup_files[0]
tumor_tmp_mpileup = cleanup_files[1]
indel_file = base + ".indel.vcf"
snp_file = base + ".snp.vcf"
cleanup_files.append(indel_file)
cleanup_files.append(snp_file)
with file_transaction(config, indel_file,
snp_file) as (tx_indel, tx_snp):
with tx_tmpdir(items[0]) as tmp_dir:
jvm_opts = _get_varscan_opts(config, tmp_dir)
fix_ambig = vcfutils.fix_ambiguous_cl()
tx_snp_in = "%s-orig" % os.path.splitext(tx_snp)[0]
tx_indel_in = "%s-orig" % os.path.splitext(tx_indel)[0]
varscan_cmd = (
"java {jvm_opts} -jar {varscan_jar} somatic"
" {normal_tmp_mpileup} {tumor_tmp_mpileup} "
"--output-snp {tx_snp_in} --output-indel {tx_indel_in} "
" --output-vcf --min-coverage 5 --p-value 0.98 "
"--strand-filter 1 ")
# add minimum AF
if "--min-var-freq" not in varscan_cmd:
min_af = float(
utils.get_in(paired.tumor_config,
("algorithm", "min_allele_fraction"),
10)) / 100.0
varscan_cmd += "--min-var-freq {min_af} "
do.run(varscan_cmd.format(**locals()), "Varscan", None, None)
for orig_fname, fname in [(tx_snp_in, tx_snp),
(tx_indel_in, tx_indel)]:
cmd = "vcfuniqalleles {orig_fname}.vcf | {fix_ambig} > {fname}"
do.run(cmd.format(**locals()), "Varscan paired fix")
# VarScan files need to be corrected to match the VCF specification
# We do this before combining them otherwise merging may fail
# if there are invalid records
to_combine = []
if do.file_exists(snp_file):
to_combine.append(snp_file)
_fix_varscan_vcf(snp_file, paired.normal_name, paired.tumor_name,
config)
if do.file_exists(indel_file):
to_combine.append(indel_file)
_fix_varscan_vcf(indel_file, paired.normal_name, paired.tumor_name,
config)
if not to_combine:
write_empty_vcf(orig_out_file, config)
return
out_file = combine_variant_files([snp_file, indel_file],
out_file,
ref_file,
config,
region=target_regions)
# Remove cleanup files
for extra_file in cleanup_files:
for ext in ["", ".gz", ".gz.tbi"]:
if os.path.exists(extra_file + ext):
os.remove(extra_file + ext)
if os.path.getsize(out_file) == 0:
write_empty_vcf(out_file)
if orig_out_file.endswith(".gz"):
out_file = bgzip_and_index(out_file, config)
_add_reject_flag(out_file, config)