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pindel_filter.v0.5.pl
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#!/usr/bin/env perl
#--------------------------------------
# @name GenomeVIP Pindel filters
# @author Beifang Niu
# @author R. Jay Mashl <[email protected]>
#
# @version 0.5 (rjm): improve documentation
# @version 0.4 (rjm): (optional) user-specified log file for appending; mode not required to be specified when not performing filtering
# @version 0.3 (rjm): generalize filter hierarchy filename handling and simply code structure
# @version 0.2 (rjm): added coverage, germline, trio, minimal pool filtering; revised approach; added failed pass. Adjusted filename and parameters names; allow for commented lines
# @version 0.1 (bn): original somatic filter, written for (tumor,normal) column order
# updated by song cao to fix the bug about the somatic part#
#--------------------------------------
use strict;
use warnings;
use Cwd;
use Carp;
use FileHandle;
use IO::File;
use Getopt::Long;
use POSIX qw( WIFEXITED );
use File::Temp qw/ tempfile /;
#my $zero=0.001;
## song changes 0.001 to 0.01 ##
my $zero=0.01;
my $thisdir;
$thisdir=`dirname $ARGV[0]`;
chomp $thisdir;
# get paras from config file
my %paras;
map { chomp; if ( !/^[#;]/ && /=/) { @_ = split /=/; $_[1] =~ s/ //g; my $v = $_[1]; $_[0] =~ s/ //g; $paras{ (split /\./, $_[0])[-1] } = $v } } (<>);
#map { print; print "\t"; print $paras{$_}; print "\n" } keys %paras;
if( $paras{'apply_filter'} eq "true" && !exists($paras{'mode'}) ) { die "Could not detect a filtering mode for filtering !!!\n"; }
# file exist ?
unless ( -e $paras{'variants_file'} ) { die "input indels not exist !!! \n"; }
my $var_file = $paras{'variants_file'};
# Filters for coverages, vaf, and balanced reads
my %filter1_prefix;
my %filter2_prefix;
$filter1_prefix{'pass'} = "";
$filter1_prefix{'fail'} = "";
$filter2_prefix{'pass'} = "";
$filter2_prefix{'fail'} = "";
if ($paras{'apply_filter'} eq "true") {
$filter1_prefix{'pass'} = "CvgVafStrand_pass";
$filter1_prefix{'fail'} = "CvgVafStrand_fail";
$filter2_prefix{'pass'} = "Homopolymer_pass";
$filter2_prefix{'fail'} = "Homopolymer_fail";
}
my $filter1_pass_fh;
my $filter1_fail_fh;
# Conversion to VCF and homopolymer filtering
my $input_fh;
my $filter2_fh_pass;
my $filter2_fh_fail;
# Optional filter, part 1
if ($paras{'apply_filter'} eq "true" && $paras{'mode'} ne "pooled") {
$input_fh = IO::File->new( "$var_file" ) or die "Could not open $var_file for reading $! ";
$filter1_pass_fh = IO::File->new( "$var_file.$filter1_prefix{'pass'}", ">" ) or die "Could not create $var_file.$filter1_prefix{'pass'} for writing $!";
$filter1_fail_fh = IO::File->new( "$var_file.$filter1_prefix{'fail'}", ">" ) or die "Could not create $var_file.$filter1_prefix{'fail'} for writing $!";
# Pindel column output
# Ref: http://seqanswers.com/forums/showthread.php?t=41121
# Ref: http://gmt.genome.wustl.edu/packages/pindel/user-manual.html
#
# In terms of 0-based numbering for three samples:
#SampleID RefSupportingLeft RefSupportingRight AltSupportingLeft AltSupportingLeftUnique AltSupportingRight AltSupportingRightUnique
# 31 32 33 34 35 36 37
# 38 39 40 41 42 43 44
# 45 46 47 48 49 50 51
# Pindel's genotyping code takes the total reference support as max($t[32],$t[33]), etc. It may be that pindel is counting
# lefts and rights separately so as to avoid misrepresenting total depth at a given location where left and right read fragments
# happen to overlap at that location. In the VAF calculations below, we implicity use ref support = avg($t[32],$t[33]).
# Germline filtering options:
# minimum coverage, VAF threshold, reads are considered balanced as long as there is nonzero read support in both directions
if ($paras{'mode'} eq "germline") {
while (<$input_fh>) {
chomp;
my @t = split /\s+/;
if( ($t[32] + $t[34] + $t[36] < $paras{'min_coverages'}) || ($t[33] + $t[34] + $t[36] < $paras{'min_coverages'}) ) {
$filter1_fail_fh->print($_."\n");
next;
}
if( ($t[34] + $t[36] + $t[34] + $t[36])/($t[32] + $t[33] + $t[34] + $t[36] + $t[34] + $t[36] ) < $paras{'min_var_allele_freq'} ){
$filter1_fail_fh->print($_."\n");
next;
}
if($paras{'require_balanced_reads'} =~ /true/) {
if ( $t[34] == 0 || $t[36] == 0 ) {
$filter1_fail_fh->print($_."\n");
next;
}
}
$filter1_pass_fh->print($_."\n");
}
}
# Somatic filtering options:
# This calculation assumes sample column order is tumor/normal, as done in GenomeVIP.
# minimum coverage met for both samples; VAF threshold in tumor; zero variant support in normal; reads are considered balanced as long as there is nonzero read support in both directions in tumor
if ($paras{'mode'} eq "somatic") {
while (<$input_fh>) {
chomp;
my @t = split /\s+/;
if( ($t[32] + $t[34] + $t[36] < $paras{'min_coverages'}) || ($t[33] + $t[34] + $t[36] < $paras{'min_coverages'}) || ($t[39] + $t[41] + $t[43] < $paras{'min_coverages'}) || ($t[40] + $t[41] + $t[43] < $paras{'min_coverages'})) {
$filter1_fail_fh->print($_."\n");
next;
}
### song changed ##
if( ($t[34] + $t[36] + $t[34] + $t[36])/($t[32] + $t[33] + $t[34] + $t[36] + $t[34] + $t[36] ) > $zero || ($t[41] + $t[43] + $t[41] + $t[43])/($t[39] + $t[40] + $t[41] + $t[43] + $t[41] + $t[43]) < $paras{'min_var_allele_freq'}) {
# if( ($t[34] + $t[36] + $t[34] + $t[36])/($t[32] + $t[33] + $t[34] + $t[36] + $t[34] + $t[36] ) < $paras{'min_var_allele_freq'} || ($t[41] + $t[43] + $t[41] + $t[43])/($t[39] + $t[40] + $t[41] + $t[43] + $t[41] + $t[43]) > $zero) {
$filter1_fail_fh->print($_."\n");
next;
}
if($paras{'require_balanced_reads'} =~ /true/) {
if ( $t[41] == 0 || $t[43] == 0 ) {
#if ( $t[34] == 0 || $t[36] == 0 || $t[41] > 0 || $t[43] > 0 ) {
$filter1_fail_fh->print($_."\n");
next;
}
}
if($paras{'remove_complex_indels'} =~ /true/) {
if ( $t[1] eq "I" || $t[1] eq "D") {
if ( $t[1] eq "I" || ($t[1] eq "D" && $t[4] == 0) ) {
print "Indel filter: passed\n";
$filter1_pass_fh->print($_."\n");
} else {
$filter1_fail_fh->print($_."\n");
}
next;
}
}
$filter1_pass_fh->print($_."\n");
}
}
# Trio filtering options:
# This calculation assumes sample column order is parent/parent/child, as done in GenomeVIP.
# minimum coverage met for all samples; VAF threshold in child; maximum allowed variant support in parents combined; reads are considered balanced as long as there is nonzero read support in both directions in child
if ($paras{'mode'} eq "trio") { # trio
while (<$input_fh>) {
chomp;
my @t = split /\s+/;
if( ($t[32] + $t[34] + $t[36] < $paras{'min_coverages'}) || ($t[33] + $t[34] + $t[36] < $paras{'min_coverages'}) || ($t[39] + $t[41] + $t[43] < $paras{'min_coverages'}) || ($t[40] + $t[41] + $t[43] < $paras{'min_coverages'}) || ($t[46] + $t[48] + $t[50] < $paras{'min_coverages'}) || ($t[47] + $t[48] + $t[50] < $paras{'min_coverages'}) ) {
$filter1_fail_fh->print($_."\n");
next;
}
if( ($t[48] + $t[50] + $t[48] + $t[50])/($t[46] + $t[47] + $t[48] + $t[50] + $t[48] + $t[50]) < $paras{'child_var_allele_freq'} ) {
$filter1_fail_fh->print($_."\n");
next;
}
if( ($t[34] + $t[36]) + ($t[41] + $t[43]) > $paras{'parents_max_num_supporting_reads'} ) {
$filter1_fail_fh->print($_."\n");
next;
}
if($paras{'require_balanced_reads'} =~ /true/) {
if ( $t[48] == 0 || $t[50] == 0 ) {
$filter1_fail_fh->print($_."\n");
next;
}
}
$filter1_pass_fh->print($_."\n");
}
}
$filter1_fail_fh->close;
$filter1_pass_fh->close;
$input_fh->close;
}
# run pindel2vcf
my $pindel2vcf_command = "";
my $ref_base = `basename $paras{'REF'}`;
chomp $ref_base;
my $result;
my $var_src;
my $var_dest;
if ($paras{'apply_filter'} eq "true" && $paras{'mode'} ne "pooled") { # only case for filter1
$var_src = "$var_file.$filter1_prefix{'pass'}";
$var_dest = "$var_file.$filter1_prefix{'pass'}.vcf";
} else {
$var_src = "$var_file";
$var_dest = "$var_file.vcf";
}
my $logfile="pindel2vcf.log";
if(exists($paras{'logfile'})) { $logfile=$paras{'logfile'}; }
# NOTE: pindel -co option seems to work on vcf output but warning messages may happend regardless
$pindel2vcf_command = "$paras{'pindel2vcf'} -r $paras{'REF'} -R $ref_base -p $var_src -d $paras{'date'} -v $var_dest -he $paras{'heterozyg_min_var_allele_freq'} -ho $paras{'homozyg_min_var_allele_freq'} >> $thisdir/$logfile 2>&1";
# print $pindel2vcf_command."\n";
$result = system( $pindel2vcf_command );
# Optional filter, part 2; here pooled is ok
if ($paras{'apply_filter'} eq "true") {
if ($paras{'mode'} eq "pooled") {
$input_fh = IO::File->new( "$var_file.vcf" ) or die "Could not open $var_file.vcf for reading $!";
$filter2_fh_pass = IO::File->new( "$var_file.$filter2_prefix{'pass'}.vcf", ">") or die "Could not create $var_file.$filter2_prefix{'pass'}.vcf for writing $!";
$filter2_fh_fail = IO::File->new( "$var_file.$filter2_prefix{'fail'}.vcf", ">") or die "Could not create $var_file.$filter2_prefix{'fail'}.vcf for writing $!";
} else {
$input_fh = IO::File->new( "$var_file.$filter1_prefix{'pass'}.vcf" ) or die "Could not open $var_file.$filter1_prefix{'pass'}.vcf for reading $!";
$filter2_fh_pass = IO::File->new( "$var_file.$filter1_prefix{'pass'}.$filter2_prefix{'pass'}.vcf", ">") or die "Could not create $var_file.$filter1_prefix{'pass'}.$filter2_prefix{'pass'}.vcf for writing $!";
$filter2_fh_fail = IO::File->new( "$var_file.$filter1_prefix{'pass'}.$filter2_prefix{'fail'}.vcf", ">") or die "Could not create $var_file.$filter1_prefix{'pass'}.$filter2_prefix{'fail'}.vcf for writing $!";
}
while ( <$input_fh> ) {
# print;
if ( /^#/ ) { $filter2_fh_pass->print($_); next };
my @a= split /\t/;
my @b = split/\;/, $a[7];
for ( my $i=0; $i<scalar(@b); $i++) {
if ( $b[$i]=~/^HOMLEN/ ) {
my @c = split/=/, $b[$i];
if ( $c[1] <= $paras{'max_num_homopolymer_repeat_units'} ) {
$filter2_fh_pass->print($_);
} else {
$filter2_fh_fail->print($_);
}
last;
}
}
}
$filter2_fh_fail->close;
$filter2_fh_pass->close;
$input_fh->close;
}
1;