13 &usage if (@ARGV < 1);
14 my $command = shift(@ARGV);
15 my %func = (subsam=>\&subsam, listsam=>\&listsam, fillac=>\&fillac, qstats=>\&qstats, varFilter=>\&varFilter,
16 hapmap2vcf=>\&hapmap2vcf, ucscsnp2vcf=>\&ucscsnp2vcf, filter4vcf=>\&varFilter, ldstats=>\&ldstats,
17 gapstats=>\&gapstats, splitchr=>\&splitchr, vcf2fq=>\&vcf2fq);
18 die("Unknown command \"$command\".\n") if (!defined($func{$command}));
23 my %opts = (l=>5000000);
24 getopts('l:', \%opts);
26 die(qq/Usage: vcfutils.pl splitchr [-l $opts{l}] <in.fa.fai>\n/) if (@ARGV == 0 && -t STDIN);
30 for (my $i = 0; $i < $t[1];) {
31 my $e = ($t[1] - $i) / $l < 1.1? $t[1] : $i + $l;
32 print "$t[0]:".($i+1)."-$e\n";
39 die(qq/Usage: vcfutils.pl subsam <in.vcf> [samples]\n/) if (@ARGV == 0);
41 my $fn = shift(@ARGV);
43 open($fh, ($fn =~ /\.gz$/)? "gzip -dc $fn |" : $fn) || die;
44 $h{$_} = 1 for (@ARGV);
50 my @s = @t[0..8]; # all fixed fields + FORMAT
57 pop(@s) if (@s == 9); # no sample selected; remove the FORMAT field
58 print join("\t", @s), "\n";
62 print join("\t", @t[0..7]), "\n";
64 print join("\t", @t[0..8], map {$t[$_]} @col), "\n";
72 die(qq/Usage: vcfutils.pl listsam <in.vcf>\n/) if (@ARGV == 0 && -t STDIN);
76 print join("\n", @t[9..$#t]), "\n";
83 die(qq/Usage: vcfutils.pl fillac <in.vcf>\n\nNote: The GT field MUST BE present and always appear as the first field.\n/) if (@ARGV == 0 && -t STDIN);
92 @_ = split(":", $t[8]);
94 if ($_[$_] eq 'GT') { $s = $_; last; }
97 print join("\t", @t), "\n";
101 if ($t[$_] =~ /^0,0,0/) {
102 } elsif ($t[$_] =~ /^([^\s:]+:){$s}(\d+).(\d+)/) {
107 my $AC = "AC=" . join("\t", @c[1..$#c]) . ";AN=$n";
109 $info =~ s/(;?)AC=(\d+)//;
110 $info =~ s/(;?)AN=(\d+)//;
117 print join("\t", @t), "\n";
124 getopts('t:', \%opts);
125 die("Usage: vcfutils.pl ldstats [-t $opts{t}] <in.vcf>\n") if (@ARGV == 0 && -t STDIN);
126 my $cutoff = $opts{t};
127 my ($last, $lastchr) = (0x7fffffff, '');
128 my ($x, $y, $n) = (0, 0, 0);
130 if (/^([^#\s]+)\s(\d+)/) {
131 my ($chr, $pos) = ($1, $2);
132 if (/NEIR=([\d\.]+)/) {
134 ++$y, $x += $pos - $last if ($lastchr eq $chr && $pos > $last && $1 > $cutoff);
136 $last = $pos; $lastchr = $chr;
139 print "Number of SNP intervals in strong LD (r > $opts{t}): $y\n";
140 print "Fraction: ", $y/$n, "\n";
141 print "Length: $x\n";
145 my %opts = (r=>'', s=>0.02, v=>undef);
146 getopts('r:s:v', \%opts);
147 die("Usage: vcfutils.pl qstats [-r ref.vcf] <in.vcf>\n
148 Note: This command discards indels. Output: QUAL #non-indel #SNPs #transitions #joint ts/tv #joint/#ref #joint/#non-indel \n") if (@ARGV == 0 && -t STDIN);
149 my %ts = (AG=>1, GA=>1, CT=>1, TC=>1);
151 my $is_vcf = defined($opts{v})? 1 : 0;
152 if ($opts{r}) { # read the reference positions
154 open($fh, $opts{r}) || die;
159 $h{$t[0],$t[1]} = $t[4];
161 $h{$1,$2} = 1 if (/^(\S+)\s+(\d+)/);
166 my $hsize = scalar(keys %h);
171 next if (length($t[3]) != 1 || uc($t[3]) eq 'N');
172 $t[3] = uc($t[3]); $t[4] = uc($t[4]);
173 my @s = split(',', $t[4]);
174 $t[5] = 3 if ($t[5] eq '.' || $t[5] < 0);
175 next if (length($s[0]) != 1);
179 my $aa = $h{$t[0],$t[1]};
181 my @aaa = split(",", $aa);
183 $hit = 1 if ($_ eq $s[0]);
187 $hit = defined($h{$t[0],$t[1]})? 1 : 0;
189 push(@a, [$t[5], ($t[4] eq '.' || $t[4] eq $t[3])? 0 : 1, $ts{$t[3].$s[0]}? 1 : 0, $hit]);
191 push(@a, [-1, 0, 0, 0]); # end marker
192 die("[qstats] No SNP data!\n") if (@a == 0);
193 @a = sort {$b->[0]<=>$a->[0]} @a;
196 my @c = (0, 0, 0, 0);
200 if ($p->[0] == -1 || ($p->[0] != $last && $c[0]/@a > $next)) {
202 $x[0] = sprintf("%.4f", $c[1]-$c[2]? $c[2] / ($c[1] - $c[2]) : 100);
203 $x[1] = sprintf("%.4f", $hsize? $c[3] / $hsize : 0);
204 $x[2] = sprintf("%.4f", $c[3] / $c[1]);
205 my $a = $c[1] - $lc[1];
206 my $b = $c[2] - $lc[2];
207 $x[3] = sprintf("%.4f", $a-$b? $b / ($a-$b) : 100);
208 print join("\t", $last, @c, @x), "\n";
209 $next = $c[0]/@a + $opts{s};
210 $lc[1] = $c[1]; $lc[2] = $c[2];
212 ++$c[0]; $c[1] += $p->[1]; $c[2] += $p->[2]; $c[3] += $p->[3];
218 my %opts = (d=>2, D=>10000, a=>2, W=>10, Q=>10, w=>10, p=>undef, 1=>1e-4, 2=>1e-100, 3=>0, 4=>1e-4, G=>0, S=>1000);
219 getopts('pd:D:W:Q:w:a:1:2:3:4:G:S:', \%opts);
221 Usage: vcfutils.pl varFilter [options] <in.vcf>
223 Options: -Q INT minimum RMS mapping quality for SNPs [$opts{Q}]
224 -d INT minimum read depth [$opts{d}]
225 -D INT maximum read depth [$opts{D}]
226 -a INT minimum number of alternate bases [$opts{a}]
227 -w INT SNP within INT bp around a gap to be filtered [$opts{w}]
228 -W INT window size for filtering adjacent gaps [$opts{W}]
229 -1 FLOAT min P-value for strand bias (given PV4) [$opts{1}]
230 -2 FLOAT min P-value for baseQ bias [$opts{2}]
231 -3 FLOAT min P-value for mapQ bias [$opts{3}]
232 -4 FLOAT min P-value for end distance bias [$opts{4}]
233 -p print filtered variants
235 Note: Some of the filters rely on annotations generated by SAMtools\/BCFtools.
236 \n/) if (@ARGV == 0 && -t STDIN);
238 # calculate the window size
239 my ($ol, $ow) = ($opts{W}, $opts{w});
240 my $max_dist = $ol > $ow? $ol : $ow;
242 my @staging; # (indel_filtering_score, flt_tag, indel_span; chr, pos, ...)
248 next if ($t[4] eq '.'); # skip non-var sites
249 # check if the site is a SNP
251 if (length($t[3]) > 1) {
253 my @s = split(',', $t[4]);
255 $type = 3 if (length != length($t[3]));
258 my @s = split(',', $t[4]);
260 $type = 3 if (length > 1);
263 # clear the out-of-range elements
265 # Still on the same chromosome and the first element's window still affects this position?
266 last if ($staging[0][3] eq $t[0] && $staging[0][4] + $staging[0][2] + $max_dist >= $t[1]);
267 varFilter_aux(shift(@staging), $opts{p}); # calling a function is a bit slower, not much
271 my ($dp, $mq, $dp_alt) = (-1, -1, -1);
272 if ($t[7] =~ /DP4=(\d+),(\d+),(\d+),(\d+)/i) {
273 $dp = $1 + $2 + $3 + $4;
276 if ($t[7] =~ /DP=(\d+)/i) {
279 $mq = $1 if ($t[7] =~ /MQ=(\d+)/i);
280 # the depth and mapQ filter
282 if ($dp < $opts{d}) {
284 } elsif ($dp > $opts{D}) {
288 $flt = 4 if ($dp_alt >= 0 && $dp_alt < $opts{a});
289 $flt = 1 if ($flt == 0 && $mq >= 0 && $mq < $opts{Q});
290 $flt = 7 if ($flt == 0 && /PV4=([^,]+),([^,]+),([^,]+),([^,;\t]+)/
291 && ($1<$opts{1} || $2<$opts{2} || $3<$opts{3} || $4<$opts{4}));
292 $flt = 8 if ($flt == 0 && ((/MXGQ=(\d+)/ && $1 >= $opts{G}) || (/MXSP=(\d+)/ && $1 < $opts{S})));
294 my $score = $t[5] * 100 + $dp_alt;
295 my $rlen = length($t[3]) - 1; # $indel_score<0 for SNPs
297 if ($type == 3) { # an indel
298 # filtering SNPs and MNPs
299 for my $x (@staging) {
300 next if (($x->[0]&3) == 3 || $x->[1] || $x->[4] + $x->[2] + $ow < $t[1]);
303 # check the staging list for indel filtering
304 for my $x (@staging) {
305 next if (($x->[0]&3) != 3 || $x->[1] || $x->[4] + $x->[2] + $ol < $t[1]);
306 if ($x->[0]>>2 < $score) {
312 } else { # SNP or MNP
313 for my $x (@staging) {
314 next if (($x->[0]&3) != 3 || $x->[4] + $x->[2] + $ow < $t[1]);
319 for my $x (@staging) {
320 next if (($x->[0]&3) == 3 || $x->[4] + $x->[2] < $t[1]);
321 if ($x->[0]>>2 < $score) {
329 push(@staging, [$score<<2|$type, $flt, $rlen, @t]);
331 # output the last few elements in the staging list
333 varFilter_aux(shift @staging, $opts{p});
338 my ($first, $is_print) = @_;
339 if ($first->[1] == 0) {
340 print join("\t", @$first[3 .. @$first-1]), "\n";
341 } elsif ($is_print) {
342 print STDERR join("\t", substr("UQdDaGgPMS", $first->[1], 1), @$first[3 .. @$first-1]), "\n";
348 $c0[$_] = $c1[$_] = 0 for (0 .. 10000);
352 next if (length($t[3]) == 1 && $t[4] =~ /^[A-Za-z](,[A-Za-z])*$/); # not an indel
353 my @s = split(',', $t[4]);
355 my $l = length($x) - length($t[3]) + 5000;
357 $l = -(length($x) - 1) + 5000;
358 } elsif ($x =~ /^\+/) {
359 $l = length($x) - 1 + 5000;
364 for (my $i = 0; $i < 10000; ++$i) {
365 next if ($c0[$i] == 0);
367 $c1[1] += $c0[$i] if (($i-5000)%3 == 0);
368 printf("C\t%d\t%.2f\n", ($i-5000), $c0[$i]);
370 printf("3\t%d\t%d\t%.3f\n", $c1[0], $c1[1], $c1[1]/$c1[0]);
374 die("Usage: vcfutils.pl <in.ucsc.snp>\n") if (@ARGV == 0 && -t STDIN);
375 print "##fileformat=VCFv4.0\n";
376 print join("\t", "#CHROM\tPOS\tID\tREF\tALT\tQUAL\tFILTER\tINFO"), "\n";
379 my $indel = ($t[9] =~ /^[ACGT](\/[ACGT])+$/)? 0 : 1;
384 $t[9] = reverse($t[9]);
385 $t[9] =~ tr/ACGTRYMKWSNacgtrymkwsn/TGCAYRKMWSNtgcayrkmwsn/;
387 my @a = split("/", $t[9]);
389 push(@alt, $_) if ($_ ne $alt[0]);
395 $alt[$_] = "N$alt[$_]";
398 my $ref = shift(@alt);
399 my $af = $t[13] > 0? ";AF=$t[13]" : '';
400 my $valid = ($t[12] eq 'unknown')? '' : ";valid=$t[12]";
401 my $info = "molType=$t[10];class=$t[11]$valid$af";
402 print join("\t", $t[1], $pos, $t[4], $ref, join(",", @alt), 0, '.', $info), "\n";
407 die("Usage: vcfutils.pl <in.ucsc.snp> <in.hapmap>\n") if (@ARGV == 0);
408 my $fn = shift(@ARGV);
410 warn("Parsing UCSC SNPs...\n");
412 open($fh, ($fn =~ /\.gz$/)? "gzip -dc $fn |" : $fn) || die;
415 next if ($t[3] - $t[2] != 1); # not SNP
416 @{$map{$t[4]}} = @t[1,3,7];
420 warn("Writing VCF...\n");
421 print "##fileformat=VCFv4.0\n";
424 if ($t[0] eq 'rs#') { # the first line
425 print join("\t", "#CHROM\tPOS\tID\tREF\tALT\tQUAL\tFILTER\tINFO\tFORMAT", @t[11..$#t]), "\n";
427 next unless ($map{$t[0]});
428 next if (length($t[1]) != 3); # skip non-SNPs
429 my $a = \@{$map{$t[0]}};
431 my @u = split('/', $t[1]);
433 $u[1] = $u[0]; $u[0] = $ref;
434 } elsif ($u[0] ne $ref) { next; }
437 $w{$u[0]} = 0; $w{$u[1]} = 1;
438 my @s = (@$a[0,1], $t[0], $ref, $alt, 0, '.', '.', 'GT');
444 my @a = ($w{substr($_,0,1)}, $w{substr($_,1,1)});
445 if (!defined($a[0]) || !defined($a[1])) {
449 push(@s, "$a[0]/$a[1]");
453 print join("\t", @s), "\n";
459 my %opts = (d=>3, D=>100000, Q=>10, l=>5);
460 getopts('d:D:Q:l:', \%opts);
462 Usage: vcfutils.pl vcf2fq [options] <all-site.vcf>
464 Options: -d INT minimum depth [$opts{d}]
465 -D INT maximum depth [$opts{D}]
466 -Q INT min RMS mapQ [$opts{Q}]
467 -l INT INDEL filtering window [$opts{l}]
468 \n/) if (@ARGV == 0 && -t STDIN);
470 my ($last_chr, $seq, $qual, $last_pos, @gaps);
475 my %het = (AC=>'M', AG=>'R', AT=>'W', CA=>'M', CG=>'S', CT=>'Y',
476 GA=>'R', GC=>'S', GT=>'K', TA=>'W', TC=>'Y', TG=>'K');
482 if ($last_chr ne $t[0]) {
483 &v2q_post_process($last_chr, \$seq, \$qual, \@gaps, $opts{l}) if ($last_chr);
484 ($last_chr, $last_pos) = ($t[0], 0);
488 die("[vcf2fq] unsorted input\n") if ($t[1] - $last_pos < 0);
489 if ($t[1] - $last_pos > 1) {
490 $seq .= 'n' x ($t[1] - $last_pos - 1);
491 $qual .= '!' x ($t[1] - $last_pos - 1);
493 if (length($t[3]) == 1 && $t[7] !~ /INDEL/ && $t[4] =~ /^([A-Za-z.])(,[A-Za-z])*$/) { # a SNP or reference
494 my ($ref, $alt) = ($t[3], $1);
496 $q = $1 if ($t[7] =~ /FQ=(-?[\d\.]+)/);
498 $_ = $1 if ($t[7] =~ /AF1=([\d\.]+)/);
499 $b = ($_ < .5 || $alt eq '.')? $ref : $alt;
502 $b = $het{"$ref$alt"};
506 $b = uc($b) if (($t[7] =~ /MQ=(\d+)/ && $1 >= $_Q) && ($t[7] =~ /DP=(\d+)/ && $1 >= $_d && $1 <= $_D));
507 $q = int($q + 33 + .499);
508 $q = chr($q <= 126? $q : 126);
512 push(@gaps, [$t[1], length($t[3])]);
516 &v2q_post_process($last_chr, \$seq, \$qual, \@gaps, $opts{l});
519 sub v2q_post_process {
520 my ($chr, $seq, $qual, $gaps, $l) = @_;
522 my $beg = $g->[0] > $l? $g->[0] - $l : 0;
523 my $end = $g->[0] + $g->[1] + $l;
524 $end = length($$seq) if ($end > length($$seq));
525 substr($$seq, $beg, $end - $beg) = lc(substr($$seq, $beg, $end - $beg));
527 print "\@$chr\n"; &v2q_print_str($seq);
528 print "+\n"; &v2q_print_str($qual);
534 for (my $i = 0; $i < $l; $i += 60) {
535 print substr($$s, $i, 60), "\n";
541 Usage: vcfutils.pl <command> [<arguments>]\n
542 Command: subsam get a subset of samples
543 listsam list the samples
544 fillac fill the allele count field
545 qstats SNP stats stratified by QUAL
547 hapmap2vcf convert the hapmap format to VCF
548 ucscsnp2vcf convert UCSC SNP SQL dump to VCF
550 varFilter filtering short variants (*)
551 vcf2fq VCF->fastq (**)
553 Notes: Commands with description endting with (*) may need bcftools
554 specific annotations.