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1 #!/usr/bin/env perl
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2
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3 =head1 LICENSE
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4
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5 Strelka Workflow Software
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6 Copyright (c) 2009-2013 Illumina, Inc.
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7
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8 This software is provided under the terms and conditions of the
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9 Illumina Open Source Software License 1.
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10
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11 You should have received a copy of the Illumina Open Source
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12 Software License 1 along with this program. If not, see
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13 <https://github.com/downloads/sequencing/licenses/>.
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14
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15 =head1 SYNOPSIS
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16
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17 configureStrelkaWorkflow.pl --tumor FILE --normal FILE --ref FILE --config FILE [options]
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18
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19 This script configures the strelka workflow for somatic variant
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20 calling on matched tumor-normal BAM files. The configuration process
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21 will produce an analysis makefile and directory structure. The
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22 makefile can be used to run the analysis on a workstation or compute
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23 cluster via make/qmake or other makefile compatible process.
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24
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25 =head1 ARGUMENTS
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26
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27 =over 4
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28
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29 =item --tumor FILE
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30
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31 Path to tumor sample BAM file (required)
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32
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33 =item --normal FILE
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34
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35 Path to normal sample BAM file (required)
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36
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37 =item --ref FILE
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38
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39 Path to reference genome fasta (required)
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40
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41 =item --config FILE
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42
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43 Strelka configuration file. Default config files can be found in
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44 ${STRELKA_INSTALL_ROOT}/etc/ for both ELAND and BWA alignments. (required)
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45
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46 =back
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47
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48 =head1 OPTIONS
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49
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50 =over 4
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51
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52 =item --output-dir DIRECTORY
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53
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54 Root of the analysis directory. This script will place all
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55 configuration files in the analysis directory, and after configuration
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56 all results and intermediate files will be written within the analysis
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57 directory during a run. This directory must not already
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58 exist. (default: ./strelkaAnalysis)
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59
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60 =back
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61
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62 =cut
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63
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64 use warnings FATAL => 'all';
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65 use strict;
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66
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67 use Carp;
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68 $SIG{__DIE__} = \&Carp::confess;
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69
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70 use Cwd qw(getcwd);
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71 use File::Spec;
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72 use Getopt::Long;
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73 use Pod::Usage;
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74
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75 my $baseDir;
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76 my $libDir;
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77 BEGIN {
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78 my $thisDir=(File::Spec->splitpath($0))[1];
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79 $baseDir=$thisDir;#$baseDir=File::Spec->catdir($thisDir,File::Spec->updir());
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80 $libDir=File::Spec->catdir($baseDir,'lib');
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81 }
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82 use lib $libDir;
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83 use Utils;
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84
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85 if(getAbsPath($baseDir)) {
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86 errorX("Can't resolve path for strelka_workflow install directory: '$baseDir'");
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87 }
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88 my $libexecDir=File::Spec->catdir($baseDir,'libexec');
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89 #my $optDir=File::Spec->catdir($baseDir,'opt');
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90
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91
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92 my $scriptName=(File::Spec->splitpath($0))[2];
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93 my $argCount=scalar(@ARGV);
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94 my $cmdline = join(' ',$0,@ARGV);
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95
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96
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97 sub usage() { pod2usage(-verbose => 1,
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98 -exitval => 2); }
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99
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100 #
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101 # user configuration:
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102 #
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103
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104 my ($tumorBam, $normalBam, $refFile, $configFile, $outDir);
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105 my $help;
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106
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107 GetOptions( "tumor=s" => \$tumorBam,
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108 "normal=s" => \$normalBam,
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109 "ref=s" => \$refFile,
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110 "config=s" => \$configFile,
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111 "output-dir=s" => \$outDir,
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112 "help|h" => \$help) || usage();
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113
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114 usage() if($help);
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115 usage() unless($argCount);
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116
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117
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118 #
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119 # Validate input conditions:
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120 #
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121
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122 sub checkFileArg($$) {
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123 my ($file,$label) = @_;
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124
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125 errorX("Must specify $label file") unless(defined($file)); #raise an error if file not defined
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126 checkFile($file,$label);
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127 }
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128
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129 checkFileArg($tumorBam,"tumor BAM");
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130 checkFileArg($normalBam,"normal BAM");
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131 checkFileArg($refFile,"reference fasta");
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132 checkFileArg($configFile,"configuration ini");
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133
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134 sub makeAbsoluteFilePaths(\$) {
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135 my ($filePathRef) = @_;
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136
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137 my ($v,$fileDir,$fileName) = File::Spec->splitpath($$filePathRef);
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138 if(getAbsPath($fileDir)) {
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139 errorX("Can't resolve directory path for '$fileDir' from input file argument: '$$filePathRef'");
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140 }
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141 $$filePathRef = File::Spec->catfile($fileDir,$fileName);
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142 }
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143
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144 makeAbsoluteFilePaths($tumorBam);
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145 makeAbsoluteFilePaths($normalBam);
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146 makeAbsoluteFilePaths($refFile);
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147 makeAbsoluteFilePaths($configFile);
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148
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149 # also check for BAM index files:
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150 sub checkBamIndex($) {
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151 my ($file) = @_;
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152 my $ifile = $file . ".bai";
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153 if(! -f $ifile) {
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154 errorX("Can't find index for BAM file '$file'");
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155 }
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156 }
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157
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158 checkBamIndex($tumorBam);
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159 checkBamIndex($normalBam);
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160
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161
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162 sub checkFaIndex($) {
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163 my ($file) = @_;
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164 my $ifile = $file . ".fai";
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165 if(! -f $ifile) {
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166 errorX("Can't find index for fasta file '$file'");
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167 }
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168 # check that fai file isn't improperly formatted (a la the GATK bundle NCBI 37 fai files)
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169 open(my $FH,"< $ifile") || errorX("Can't open fai file '$ifile'");
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170 my $lineno=1;
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171 while(<$FH>) {
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172 chomp;
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173 my @F=split();
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174 if(scalar(@F) != 5) {
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175 errorX("Unexpected format for line number '$lineno' of fasta index file: '$ifile'\n\tRe-running fasta indexing may fix the issue. To do so, run: \"samtools faidx $file\"");
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176 }
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177 $lineno++;
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178 }
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179 close($FH);
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180 }
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181
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182 checkFaIndex($refFile);
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183
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184
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185 if(defined($outDir)) {
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186 if(getAbsPath($outDir)) {
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187 errorX("Can't resolve path for ouput directory: '$outDir'");
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188 }
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189 } else {
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190 $outDir=File::Spec->catdir(Cwd::getcwd(),'strelkaAnalysis');
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191 }
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192
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193 if(-e $outDir) {
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194 errorX("Output path already exists: '$outDir'");
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195 }
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196
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197 if(getAbsPath($baseDir)) {
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198 errorX("Can't resolve path for strelka install directory: '$baseDir'");
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199 }
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200
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201 #my $samtoolsDir = File::Spec->catdir($optDir,'samtools');
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202 checkDir($libexecDir,"strelka libexec");
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203 #checkDir($samtoolsDir,"samtools");
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204
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205 my $callScriptName = "callSomaticVariants.pl";
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206 my $filterScriptName = "filterSomaticVariants.pl";
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207 my $finishScriptName = "consolidateResults.pl";
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208 my $callScript = File::Spec->catfile($libexecDir,$callScriptName);
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209 my $filterScript = File::Spec->catfile($libexecDir,$filterScriptName);
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210 my $finishScript = File::Spec->catfile($libexecDir,$finishScriptName);
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211 my $countFasta = File::Spec->catfile($libexecDir,"countFastaBases");
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212 #my $samtoolsBin = File::Spec->catfile($samtoolsDir,"samtools");
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213 checkFile($callScript,"somatic variant call script");
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214 checkFile($filterScript,"somatic variant filter script");
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215 checkFile($finishScript,"result consolidation script");
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216 checkFile($countFasta,"fasta scanner");
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217 #checkFile($samtoolsBin,"samtools");
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218
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219 #
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220 # Configure bin runs:
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221 #
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222 checkMakeDir($outDir);
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223
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224 #
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225 # Configure bin runs: open and validate config ini
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226 #
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227 my $config = parseConfigIni($configFile);
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228
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229 sub checkConfigKeys($) {
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230 my ($keyref) = @_;
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231 for (@$keyref) {
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232 errorX("Undefined configuration option: '$_'") unless(defined($config->{user}{$_}));
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233 }
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234 }
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235
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236 # these are the keys we need at configuration time:
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237 my @config_keys = qw(binSize);
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238
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239 # these are additional keys we will need to run the workflow:
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240 # (note we don't check for (maxInputDepth,minTier2Mapq) for back compatibility with older config files)
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241 my @workflow_keys = qw(
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242 minTier1Mapq isWriteRealignedBam ssnvPrior sindelPrior ssnvNoise sindelNoise ssnvNoiseStrandBiasFrac
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243 ssnvQuality_LowerBound sindelQuality_LowerBound isSkipDepthFilters depthFilterMultiple
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244 snvMaxFilteredBasecallFrac snvMaxSpanningDeletionFrac indelMaxRefRepeat
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245 indelMaxWindowFilteredBasecallFrac indelMaxIntHpolLength);
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246
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247 checkConfigKeys(\@config_keys);
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248 checkConfigKeys(\@workflow_keys);
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249
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250
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251 my $binSize = int($config->{user}{binSize});
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252
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253 $config->{derived}{configurationCmdline} = $cmdline;
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254 $config->{derived}{normalBam} = $normalBam;
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255 $config->{derived}{tumorBam} = $tumorBam;
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256 $config->{derived}{refFile} = $refFile;
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257 $config->{derived}{outDir} = $outDir;
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258
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259 #
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260 # Configure bin runs: check for consistent chrom info between BAMs and reference
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261 #
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262 sub getBamChromInfo($) {
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263 my $file = shift;
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264 my $cmd = "samtools view -H $file |";
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265 open(my $FH,$cmd) || errorX("Can't open process $cmd");
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266
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267 my %info;
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268 my $n=0;
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269 while(<$FH>) {
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270 next unless(/^\@SQ/);
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271 chomp;
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272 my @F = split(/\t/);
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273 scalar(@F) >= 3 || errorX("Unexpected bam header for file '$file'");
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274
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275 my %h = ();
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276 foreach (@F) {
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277 my @vals = split(':');
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278 $h{$vals[0]} = $vals[1];
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279 }
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280 $F[1] = $h{'SN'};
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281 $F[2] = $h{'LN'};
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282
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283 my $size = int($F[2]);
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284 ($size > 0) || errorX("Unexpected chromosome size '$size' in bam header for file '$file'");
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285 $info{$F[1]}{size} = $size;
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286 $info{$F[1]}{order} = $n;
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287 $n++;
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288 }
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289 close($FH) || errorX("Can't close process $cmd");
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290 return %info;
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291 }
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292
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293
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294 my %chromInfo = getBamChromInfo($normalBam);
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295 my @chroms = sort { $chromInfo{$a}{order} <=> $chromInfo{$b}{order} } (keys(%chromInfo));
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296
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297 {
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298 #consistency check:
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299 my %tumorChromInfo = getBamChromInfo($tumorBam);
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300 for my $chrom (@chroms) {
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301 my $ln = $chromInfo{$chrom}{size};
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302 my $tln = $tumorChromInfo{$chrom}{size};
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303 my $order = $chromInfo{$chrom}{order};
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304 my $torder = $tumorChromInfo{$chrom}{order};
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305 unless(defined($tln) && ($tln==$ln) && ($torder==$order)) {
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306 errorX("Tumor and normal BAM file headers disagree on chromosome: '$chrom'");
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307 }
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308 delete $tumorChromInfo{$chrom};
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309 }
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310 for my $chrom (keys(%tumorChromInfo)) {
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311 errorX("Tumor and normal BAM file headers disagree on chromosome: '$chrom'");
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312 }
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313 }
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314
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315
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316 my %refChromInfo;
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317 logX("Scanning reference genome");
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318 {
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319 my $knownGenomeSize=0;
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320 my $cmd="$countFasta $refFile |";
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321 open(my $FFH,$cmd) || errorX("Failed to open process '$cmd'");
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322
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323 while(<$FFH>) {
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324 chomp;
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325 my @F = split(/\t/);
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326 scalar(@F) == 4 || errorX("Unexpected value from '$cmd'");
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327 $knownGenomeSize += int($F[2]);
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328 $refChromInfo{$F[1]}{knownSize} = int($F[2]);
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329 $refChromInfo{$F[1]}{size} = int($F[3]);
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330 }
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331 close($FFH) || errorX("Failed to close process '$cmd'");
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332
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333 #consistency check:
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334 for my $chrom (@chroms) {
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335 my $ln = $chromInfo{$chrom}{size};
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336 my $rln = $refChromInfo{$chrom}{size};
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337 unless(defined($rln) && ($rln==$ln)) {
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338 errorX("BAM headers and reference fasta disagree on chromosome: '$chrom'");
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339 }
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340 $config->{derived}{"chrom_${chrom}_size"} = $rln;
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341 $config->{derived}{"chrom_${chrom}_knownSize"} = $refChromInfo{$chrom}{knownSize};
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342 }
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343 $config->{derived}{chromOrder} = join("\t",@chroms);
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344
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345 $config->{derived}{knownGenomeSize} = $knownGenomeSize;
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346 }
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347 logX("Scanning reference genome complete");
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348
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349
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350
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351 #
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352 # Configure bin runs: create directory structure
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353 #
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354 my $resultsDir = File::Spec->catdir($outDir,'results');
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355 checkMakeDir($resultsDir);
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356 if($config->{user}{isWriteRealignedBam}) {
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357 my $bamDir = File::Spec->catdir($outDir,'realigned');
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358 checkMakeDir($bamDir);
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359 }
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360 my $chromRootDir = File::Spec->catdir($outDir,'chromosomes');
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361 checkMakeDir($chromRootDir);
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362 for my $chrom (@chroms) {
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363 my $chromDir = File::Spec->catdir($chromRootDir,$chrom);
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364 checkMakeDir($chromDir);
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365
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366 my $chromRef = $chromInfo{$chrom};
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367 $chromRef->{dir} = $chromDir;
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368 $chromRef->{binList} = getBinList($chromRef->{size},$binSize);
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369
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370 my $binRootDir = File::Spec->catdir($chromDir,'bins');
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371 checkMakeDir($binRootDir);
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372
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373 for my $binId ( @{$chromRef->{binList}} ) {
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374 my $binDir = File::Spec->catdir($binRootDir,$binId);
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375 checkMakeDir($binDir);
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376 }
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377 }
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378
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379
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380
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381 #
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382 # write run config file:
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383 #
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384 my $runConfigFile;
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385 {
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386 my $cstr = <<END;
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387 ;
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388 ; Strelka workflow configuration file
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389 ;
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390 ; This is an automatically generated file, you probably don't want to edit it. If starting a new run,
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391 ; input configuration templates (with comments) can be found in the Strelka etc/ directory.
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392 ;
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393 END
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394
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395 $cstr .= writeConfigIni($config);
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396
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397 my $configDir = File::Spec->catdir($outDir,'config');
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398 checkMakeDir($configDir);
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399 $runConfigFile = File::Spec->catdir($configDir,'run.config.ini');
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400 open(my $FH,"> $runConfigFile") || errorX("Can't open file '$runConfigFile'");
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401 print $FH $cstr;
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402 close($FH);
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403 }
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404
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405
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406
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407 #
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408 # create makefile
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409 #
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410 my $makeFile = File::Spec->catfile($outDir,"Makefile");
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411 open(my $MAKEFH, "> $makeFile") || errorX("Can't open file: '$makeFile'");
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412
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413 my $completeFile = "task.complete";
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414
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415 print $MAKEFH <<ENDE;
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416 # This makefile was automatically generated by $scriptName
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417 #
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418 # Please do not edit.
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419
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420 script_dir := $libexecDir
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421 call_script := \$(script_dir)/$callScriptName
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422 filter_script := \$(script_dir)/$filterScriptName
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423 finish_script := \$(script_dir)/$finishScriptName
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424
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425 config_file := $runConfigFile
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426
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427 analysis_dir := $outDir
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428 results_dir := \$(analysis_dir)/results
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429
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430 ENDE
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431
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432 print $MAKEFH <<'ENDE';
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433
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434 complete_tag := task.complete
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435
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436 finish_task := $(analysis_dir)/$(complete_tag)
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437
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438 get_chrom_dir = $(analysis_dir)/chromosomes/$1
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439 get_chrom_task = $(call get_chrom_dir,$1)/$(complete_tag)
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440 get_bin_task = $(call get_chrom_dir,$1)/bins/$2/$(complete_tag)
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441
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442
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443
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444 all: $(finish_task)
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445 @$(print_success)
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446
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447
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448 define print_success
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449 echo;\
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450 echo Analysis complete. Final somatic calls can be found in $(results_dir);\
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451 echo
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452 endef
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453
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454
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455 # top level results target:
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456 #
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457 $(finish_task):
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458 $(finish_script) --config=$(config_file) && touch $@
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459
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460
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461 # chromosome targets:
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462 #
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463 ENDE
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464
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465 for my $chrom (@chroms) {
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466
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467 print $MAKEFH <<ENDE;
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468 chrom_${chrom}_task := \$(call get_chrom_task,$chrom)
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469 \$(finish_task): \$(chrom_${chrom}_task)
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470 \$(chrom_${chrom}_task):
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471 \$(filter_script) --config=\$(config_file) --chrom=$chrom && touch \$@
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472
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473 ENDE
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474
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475 }
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476
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477 print $MAKEFH <<ENDE;
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478
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479 # chromosome bin targets:
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480 #
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481 ENDE
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482
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483 for my $chrom (@chroms) {
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484 for my $bin (@{$chromInfo{$chrom}{binList}}) {
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485
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486 print $MAKEFH <<ENDE;
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487 chrom_${chrom}_bin_${bin}_task := \$(call get_bin_task,$chrom,$bin)
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488 \$(chrom_${chrom}_task): \$(chrom_${chrom}_bin_${bin}_task)
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489 \$(chrom_${chrom}_bin_${bin}_task):
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490 \$(call_script) --config=\$(config_file) --chrom=$chrom --bin=$bin && touch \$@
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491
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492 ENDE
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493
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494 }
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495 }
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496
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497
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498 # If the eval function is available, this is the way we could finish
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499 # the makefile without being so verbose but it doesn't look like qmake
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500 # understands this function.
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501
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502 =cut
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503
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504 print $MAKEFH <<ENDE;
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505
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506 chroms := @chroms
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507
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508 ENDE
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509
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510 for my $chrom (@chroms) {
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511 print $MAKEFH "${chrom}_bins := " . join(" ",@{$chromInfo{$chrom}{binList}}) . "\n";
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512 }
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513
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514 print $MAKEFH <<'ENDE';
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515
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516 define chrom_task_template
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517 chrom_$1_task := $(call get_chrom_task,$1)
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518 $(finish_task): $$(chrom_$1_task)
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519 $$(chrom_$1_task):
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520 $$(filter_script) --config=$$(config_file) --chrom=$1 && touch $$@
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521 endef
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522
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523 $(foreach c,$(chroms),$(eval $(call chrom_task_template,$c)))
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524
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525
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526 # chromosome bin targets:
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527 #
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528 define chrom_bin_task_template
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529 chrom_$1_bin_$2_task := $(call get_bin_task,$1,$2)
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530 $$(chrom_$1_task): $$(chrom_$1_bin_$2_task)
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531 $$(chrom_$1_bin_$2_task):
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532 $$(call_script) --config=$$(config_file) --chrom=$1 --bin=$2 && touch $$@
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533 endef
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534
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535 $(foreach c,$(chroms), \
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536 $(foreach b,$($c_bins),$(eval $(call chrom_bin_task_template,$c,$b))) \
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537 )
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538
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539 ENDE
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540
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541 =cut
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542
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543
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544
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545 print <<END;
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546
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547
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548 Successfully configured analysis and created makefile '$makeFile'.
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549
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550 To run the analysis locally using make, run:
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551
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552 make -C $outDir
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553
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554 ...or:
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555
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556 cd $outDir
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557 make
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558
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559 END
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560
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561 1;
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562
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563 __END__
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564
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