#include "bcf.h"
#include "prob1.h"
#include "kstring.h"
+#include "time.h"
-#include "khash.h"
-KHASH_SET_INIT_INT64(set64)
+#ifdef _WIN32
+#define srand48(x) srand(x)
+#define lrand48() rand()
+#endif
#include "kseq.h"
KSTREAM_INIT(gzFile, gzread, 16384)
#define VC_VARONLY 16
#define VC_VCFIN 32
#define VC_UNCOMP 64
-#define VC_HWE 128
#define VC_KEEPALT 256
#define VC_ACGT_ONLY 512
#define VC_QCALL 1024
#define VC_CALL_GT 2048
#define VC_ADJLD 4096
#define VC_NO_INDEL 8192
+#define VC_ANNO_MAX 16384
+#define VC_FIX_PL 32768
+#define VC_EM 0x10000
+#define VC_PAIRCALL 0x20000
+#define VC_QCNT 0x40000
typedef struct {
- int flag, prior_type, n1;
- char *fn_list, *prior_file;
- double theta, pref, indel_frac;
+ int flag, prior_type, n1, n_sub, *sublist, n_perm;
+ uint32_t *trio_aux;
+ char *prior_file, **subsam, *fn_dict;
+ uint8_t *ploidy;
+ double theta, pref, indel_frac, min_perm_p, min_smpl_frac, min_lrt;
+ void *bed;
} viewconf_t;
-khash_t(set64) *bcf_load_pos(const char *fn, bcf_hdr_t *_h)
-{
- void *str2id;
- gzFile fp;
- kstream_t *ks;
- int ret, dret, lineno = 1;
- kstring_t *str;
- khash_t(set64) *hash = 0;
-
- hash = kh_init(set64);
- str2id = bcf_build_refhash(_h);
- str = calloc(1, sizeof(kstring_t));
- fp = strcmp(fn, "-")? gzopen(fn, "r") : gzdopen(fileno(stdin), "r");
- ks = ks_init(fp);
- while (ks_getuntil(ks, 0, str, &dret) >= 0) {
- int tid = bcf_str2id(str2id, str->s);
- if (tid >= 0 && dret != '\n') {
- if (ks_getuntil(ks, 0, str, &dret) >= 0) {
- uint64_t x = (uint64_t)tid<<32 | (atoi(str->s) - 1);
- kh_put(set64, hash, x, &ret);
- } else break;
- } else fprintf(stderr, "[%s] %s is not a reference name (line %d).\n", __func__, str->s, lineno);
- if (dret != '\n') while ((dret = ks_getc(ks)) > 0 && dret != '\n');
- if (dret < 0) break;
- ++lineno;
- }
- bcf_str2id_destroy(str2id);
- ks_destroy(ks);
- gzclose(fp);
- free(str->s); free(str);
- return hash;
-}
-
-static double test_hwe(const double g[3])
-{
- extern double kf_gammaq(double p, double x);
- double fexp, chi2, f[3], n;
- int i;
- n = g[0] + g[1] + g[2];
- fexp = (2. * g[2] + g[1]) / (2. * n);
- if (fexp > 1. - 1e-10) fexp = 1. - 1e-10;
- if (fexp < 1e-10) fexp = 1e-10;
- f[0] = n * (1. - fexp) * (1. - fexp);
- f[1] = n * 2. * fexp * (1. - fexp);
- f[2] = n * fexp * fexp;
- for (i = 0, chi2 = 0.; i < 3; ++i)
- chi2 += (g[i] - f[i]) * (g[i] - f[i]) / f[i];
- return kf_gammaq(.5, chi2 / 2.);
-}
+void *bed_read(const char *fn);
+void bed_destroy(void *_h);
+int bed_overlap(const void *_h, const char *chr, int beg, int end);
typedef struct {
double p[4];
if ((p = strstr(b->info, "I16=")) == 0) return -1;
p += 4;
for (i = 0; i < 16; ++i) {
- anno[i] = strtol(p, &p, 10);
+ errno = 0; anno[i] = strtol(p, &p, 10);
if (anno[i] == 0 && (errno == EINVAL || errno == ERANGE)) return -2;
++p;
}
bcf_sync(b);
}
-static int update_bcf1(int n_smpl, bcf1_t *b, const bcf_p1aux_t *pa, const bcf_p1rst_t *pr, double pref, int flag)
+static int update_bcf1(bcf1_t *b, const bcf_p1aux_t *pa, const bcf_p1rst_t *pr, double pref, int flag, double em[10], int cons_llr, int64_t cons_gt)
{
kstring_t s;
- int is_var = (pr->p_ref < pref);
- double p_hwe, r = is_var? pr->p_ref : 1. - pr->p_ref;
+ int has_I16, is_var;
+ double fq, r;
anno16_t a;
- p_hwe = pr->g[0] >= 0.? test_hwe(pr->g) : 1.0; // only do HWE g[] is calculated
- test16(b, &a);
- rm_info(b, "I16=");
+ has_I16 = test16(b, &a) >= 0? 1 : 0;
+ rm_info(b, "I16="); // FIXME: probably this function has a bug. If I move it below, I16 will not be removed!
memset(&s, 0, sizeof(kstring_t));
kputc('\0', &s); kputs(b->ref, &s); kputc('\0', &s);
kputs(b->alt, &s); kputc('\0', &s); kputc('\0', &s);
kputs(b->info, &s);
if (b->info[0]) kputc(';', &s);
-// ksprintf(&s, "AF1=%.4lg;AFE=%.4lg;CI95=%.4lg,%.4lg", 1.-pr->f_em, 1.-pr->f_exp, pr->cil, pr->cih);
- ksprintf(&s, "AF1=%.4lg;CI95=%.4lg,%.4lg", 1.-pr->f_em, pr->cil, pr->cih);
- ksprintf(&s, ";DP4=%d,%d,%d,%d;MQ=%d", a.d[0], a.d[1], a.d[2], a.d[3], a.mq);
- if (a.is_tested) {
- if (pr->pc[0] >= 0.) ksprintf(&s, ";PC4=%lg,%lg,%lg,%lg", pr->pc[0], pr->pc[1], pr->pc[2], pr->pc[3]);
- ksprintf(&s, ";PV4=%.2lg,%.2lg,%.2lg,%.2lg", a.p[0], a.p[1], a.p[2], a.p[3]);
+ { // print EM
+ if (em[0] >= 0) ksprintf(&s, "AF1=%.4g", 1 - em[0]);
+ if (em[4] >= 0 && em[4] <= 0.05) ksprintf(&s, ";G3=%.4g,%.4g,%.4g;HWE=%.3g", em[3], em[2], em[1], em[4]);
+ if (em[5] >= 0 && em[6] >= 0) ksprintf(&s, ";AF2=%.4g,%.4g", 1 - em[5], 1 - em[6]);
+ if (em[7] >= 0) ksprintf(&s, ";LRT=%.3g", em[7]);
+ if (em[8] >= 0) ksprintf(&s, ";LRT2=%.3g", em[8]);
+ }
+ if (cons_llr > 0) {
+ ksprintf(&s, ";CLR=%d", cons_llr);
+ if (cons_gt > 0)
+ ksprintf(&s, ";UGT=%c%c%c;CGT=%c%c%c", cons_gt&0xff, cons_gt>>8&0xff, cons_gt>>16&0xff,
+ cons_gt>>32&0xff, cons_gt>>40&0xff, cons_gt>>48&0xff);
}
- if (pr->g[0] >= 0. && p_hwe <= .2)
- ksprintf(&s, ";GC=%.2lf,%.2lf,%.2lf;HWE=%.3lf", pr->g[2], pr->g[1], pr->g[0], p_hwe);
+ if (pr == 0) { // if pr is unset, return
+ kputc('\0', &s); kputs(b->fmt, &s); kputc('\0', &s);
+ free(b->str);
+ b->m_str = s.m; b->l_str = s.l; b->str = s.s;
+ bcf_sync(b);
+ return 1;
+ }
+
+ is_var = (pr->p_ref < pref);
+ r = is_var? pr->p_ref : pr->p_var;
+
+// ksprintf(&s, ";CI95=%.4g,%.4g", pr->cil, pr->cih); // FIXME: when EM is not used, ";" should be omitted!
+ ksprintf(&s, ";AC1=%d", pr->ac);
+ if (has_I16) ksprintf(&s, ";DP4=%d,%d,%d,%d;MQ=%d", a.d[0], a.d[1], a.d[2], a.d[3], a.mq);
+ fq = pr->p_ref_folded < 0.5? -4.343 * log(pr->p_ref_folded) : 4.343 * log(pr->p_var_folded);
+ if (fq < -999) fq = -999;
+ if (fq > 999) fq = 999;
+ ksprintf(&s, ";FQ=%.3g", fq);
+ if (pr->cmp[0] >= 0.) { // two sample groups
+ int i, q[3];
+ for (i = 1; i < 3; ++i) {
+ double x = pr->cmp[i] + pr->cmp[0]/2.;
+ q[i] = x == 0? 255 : (int)(-4.343 * log(x) + .499);
+ if (q[i] > 255) q[i] = 255;
+ }
+ if (pr->perm_rank >= 0) ksprintf(&s, ";PR=%d", pr->perm_rank);
+ // ksprintf(&s, ";LRT3=%.3g", pr->lrt);
+ ksprintf(&s, ";PCHI2=%.3g;PC2=%d,%d", q[1], q[2], pr->p_chi2);
+ }
+ if (has_I16 && a.is_tested) ksprintf(&s, ";PV4=%.2g,%.2g,%.2g,%.2g", a.p[0], a.p[1], a.p[2], a.p[3]);
kputc('\0', &s);
kputs(b->fmt, &s); kputc('\0', &s);
free(b->str);
b->m_str = s.m; b->l_str = s.l; b->str = s.s;
- b->qual = r < 1e-100? 99 : -4.343 * log(r);
- if (b->qual > 99) b->qual = 99;
+ b->qual = r < 1e-100? 999 : -4.343 * log(r);
+ if (b->qual > 999) b->qual = 999;
bcf_sync(b);
if (!is_var) bcf_shrink_alt(b, 1);
else if (!(flag&VC_KEEPALT))
b->m_str = s.m; b->l_str = s.l; b->str = s.s;
bcf_sync(b);
for (i = 0; i < b->n_smpl; ++i) {
- x = bcf_p1_call_gt(pa, pr->f_em, i);
+ x = bcf_p1_call_gt(pa, pr->f_exp, i);
((uint8_t*)b->gi[old_n_gi].data)[i] = (x&3) == 0? 1<<3|1 : (x&3) == 1? 1 : 0;
((uint8_t*)b->gi[old_n_gi+1].data)[i] = x>>2;
}
return is_var;
}
-double bcf_ld_freq(const bcf1_t *b0, const bcf1_t *b1, double f[4]);
+static char **read_samples(const char *fn, int *_n)
+{
+ gzFile fp;
+ kstream_t *ks;
+ kstring_t s;
+ int dret, n = 0, max = 0;
+ char **sam = 0;
+ *_n = 0;
+ s.l = s.m = 0; s.s = 0;
+ fp = gzopen(fn, "r");
+ if (fp == 0) return 0; // fail to open file
+ ks = ks_init(fp);
+ while (ks_getuntil(ks, 0, &s, &dret) >= 0) {
+ int l;
+ if (max == n) {
+ max = max? max<<1 : 4;
+ sam = realloc(sam, sizeof(void*)*max);
+ }
+ l = s.l;
+ sam[n] = malloc(s.l + 2);
+ strcpy(sam[n], s.s);
+ sam[n][l+1] = 2; // by default, diploid
+ if (dret != '\n') {
+ if (ks_getuntil(ks, 0, &s, &dret) >= 0) { // read ploidy, 1 or 2
+ int x = (int)s.s[0] - '0';
+ if (x == 1 || x == 2) sam[n][l+1] = x;
+ else fprintf(stderr, "(%s) ploidy can only be 1 or 2; assume diploid\n", __func__);
+ }
+ if (dret != '\n') ks_getuntil(ks, '\n', &s, &dret);
+ }
+ ++n;
+ }
+ ks_destroy(ks);
+ gzclose(fp);
+ free(s.s);
+ *_n = n;
+ return sam;
+}
+
+static void write_header(bcf_hdr_t *h)
+{
+ kstring_t str;
+ str.l = h->l_txt? h->l_txt - 1 : 0;
+ str.m = str.l + 1; str.s = h->txt;
+ if (!strstr(str.s, "##INFO=<ID=DP,"))
+ kputs("##INFO=<ID=DP,Number=1,Type=Integer,Description=\"Raw read depth\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=DP4,"))
+ kputs("##INFO=<ID=DP4,Number=4,Type=Integer,Description=\"# high-quality ref-forward bases, ref-reverse, alt-forward and alt-reverse bases\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=MQ,"))
+ kputs("##INFO=<ID=MQ,Number=1,Type=Integer,Description=\"Root-mean-square mapping quality of covering reads\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=FQ,"))
+ kputs("##INFO=<ID=FQ,Number=1,Type=Float,Description=\"Phred probability of all samples being the same\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=AF1,"))
+ kputs("##INFO=<ID=AF1,Number=1,Type=Float,Description=\"Max-likelihood estimate of the first ALT allele frequency (assuming HWE)\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=AC1,"))
+ kputs("##INFO=<ID=AC1,Number=1,Type=Float,Description=\"Max-likelihood estimate of the first ALT allele count (no HWE assumption)\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=G3,"))
+ kputs("##INFO=<ID=G3,Number=3,Type=Float,Description=\"ML estimate of genotype frequencies\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=HWE,"))
+ kputs("##INFO=<ID=HWE,Number=1,Type=Float,Description=\"Chi^2 based HWE test P-value based on G3\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=CLR,"))
+ kputs("##INFO=<ID=CLR,Number=1,Type=Integer,Description=\"Log ratio of genotype likelihoods with and without the constraint\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=UGT,"))
+ kputs("##INFO=<ID=UGT,Number=1,Type=String,Description=\"The most probable unconstrained genotype configuration in the trio\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=CGT,"))
+ kputs("##INFO=<ID=CGT,Number=1,Type=String,Description=\"The most probable constrained genotype configuration in the trio\">\n", &str);
+// if (!strstr(str.s, "##INFO=<ID=CI95,"))
+// kputs("##INFO=<ID=CI95,Number=2,Type=Float,Description=\"Equal-tail Bayesian credible interval of the site allele frequency at the 95% level\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=PV4,"))
+ kputs("##INFO=<ID=PV4,Number=4,Type=Float,Description=\"P-values for strand bias, baseQ bias, mapQ bias and tail distance bias\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=INDEL,"))
+ kputs("##INFO=<ID=INDEL,Number=0,Type=Flag,Description=\"Indicates that the variant is an INDEL.\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=PC2,"))
+ kputs("##INFO=<ID=PC2,Number=2,Type=Integer,Description=\"Phred probability of the nonRef allele frequency in group1 samples being larger (,smaller) than in group2.\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=PCHI2,"))
+ kputs("##INFO=<ID=PCHI2,Number=1,Type=Float,Description=\"Posterior weighted chi^2 P-value for testing the association between group1 and group2 samples.\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=QCHI2,"))
+ kputs("##INFO=<ID=QCHI2,Number=1,Type=Integer,Description=\"Phred scaled PCHI2.\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=RP,"))
+ kputs("##INFO=<ID=PR,Number=1,Type=Integer,Description=\"# permutations yielding a smaller PCHI2.\">\n", &str);
+ if (!strstr(str.s, "##INFO=<ID=VDB,"))
+ kputs("##INFO=<ID=VDB,Number=1,Type=Float,Description=\"Variant Distance Bias\">\n", &str);
+ if (!strstr(str.s, "##FORMAT=<ID=GT,"))
+ kputs("##FORMAT=<ID=GT,Number=1,Type=String,Description=\"Genotype\">\n", &str);
+ if (!strstr(str.s, "##FORMAT=<ID=GQ,"))
+ kputs("##FORMAT=<ID=GQ,Number=1,Type=Integer,Description=\"Genotype Quality\">\n", &str);
+ if (!strstr(str.s, "##FORMAT=<ID=GL,"))
+ kputs("##FORMAT=<ID=GL,Number=3,Type=Float,Description=\"Likelihoods for RR,RA,AA genotypes (R=ref,A=alt)\">\n", &str);
+ if (!strstr(str.s, "##FORMAT=<ID=DP,"))
+ kputs("##FORMAT=<ID=DP,Number=1,Type=Integer,Description=\"# high-quality bases\">\n", &str);
+ if (!strstr(str.s, "##FORMAT=<ID=SP,"))
+ kputs("##FORMAT=<ID=SP,Number=1,Type=Integer,Description=\"Phred-scaled strand bias P-value\">\n", &str);
+ if (!strstr(str.s, "##FORMAT=<ID=PL,"))
+ kputs("##FORMAT=<ID=PL,Number=G,Type=Integer,Description=\"List of Phred-scaled genotype likelihoods\">\n", &str);
+ h->l_txt = str.l + 1; h->txt = str.s;
+}
+
+double bcf_pair_freq(const bcf1_t *b0, const bcf1_t *b1, double f[4]);
int bcfview(int argc, char *argv[])
{
extern int bcf_2qcall(bcf_hdr_t *h, bcf1_t *b);
extern void bcf_p1_indel_prior(bcf_p1aux_t *ma, double x);
+ extern int bcf_fix_gt(bcf1_t *b);
+ extern int bcf_anno_max(bcf1_t *b);
+ extern int bcf_shuffle(bcf1_t *b, int seed);
+ extern uint32_t *bcf_trio_prep(int is_x, int is_son);
+ extern int bcf_trio_call(uint32_t *prep, const bcf1_t *b, int *llr, int64_t *gt);
+ extern int bcf_pair_call(const bcf1_t *b);
+ extern int bcf_min_diff(const bcf1_t *b);
+
bcf_t *bp, *bout = 0;
bcf1_t *b, *blast;
- int c;
- uint64_t n_processed = 0;
+ int c, *seeds = 0;
+ uint64_t n_processed = 0, qcnt[256];
viewconf_t vc;
bcf_p1aux_t *p1 = 0;
- bcf_hdr_t *h;
+ bcf_hdr_t *hin, *hout;
int tid, begin, end;
char moder[4], modew[4];
- khash_t(set64) *hash = 0;
tid = begin = end = -1;
memset(&vc, 0, sizeof(viewconf_t));
- vc.prior_type = vc.n1 = -1; vc.theta = 1e-3; vc.pref = 0.5; vc.indel_frac = -1.;
- while ((c = getopt(argc, argv, "N1:l:cHAGvbSuP:t:p:QgLi:I")) >= 0) {
+ vc.prior_type = vc.n1 = -1; vc.theta = 1e-3; vc.pref = 0.5; vc.indel_frac = -1.; vc.n_perm = 0; vc.min_perm_p = 0.01; vc.min_smpl_frac = 0; vc.min_lrt = 1;
+ memset(qcnt, 0, 8 * 256);
+ while ((c = getopt(argc, argv, "FN1:l:cC:eHAGvbSuP:t:p:QgLi:IMs:D:U:X:d:T:Y")) >= 0) {
switch (c) {
case '1': vc.n1 = atoi(optarg); break;
- case 'l': vc.fn_list = strdup(optarg); break;
+ case 'l': vc.bed = bed_read(optarg); break;
+ case 'D': vc.fn_dict = strdup(optarg); break;
+ case 'F': vc.flag |= VC_FIX_PL; break;
case 'N': vc.flag |= VC_ACGT_ONLY; break;
case 'G': vc.flag |= VC_NO_GENO; break;
case 'A': vc.flag |= VC_KEEPALT; break;
case 'b': vc.flag |= VC_BCFOUT; break;
case 'S': vc.flag |= VC_VCFIN; break;
case 'c': vc.flag |= VC_CALL; break;
+ case 'e': vc.flag |= VC_EM; break;
case 'v': vc.flag |= VC_VARONLY | VC_CALL; break;
case 'u': vc.flag |= VC_UNCOMP | VC_BCFOUT; break;
- case 'H': vc.flag |= VC_HWE; break;
case 'g': vc.flag |= VC_CALL_GT | VC_CALL; break;
case 'I': vc.flag |= VC_NO_INDEL; break;
+ case 'M': vc.flag |= VC_ANNO_MAX; break;
+ case 'Y': vc.flag |= VC_QCNT; break;
case 't': vc.theta = atof(optarg); break;
case 'p': vc.pref = atof(optarg); break;
case 'i': vc.indel_frac = atof(optarg); break;
case 'Q': vc.flag |= VC_QCALL; break;
case 'L': vc.flag |= VC_ADJLD; break;
+ case 'U': vc.n_perm = atoi(optarg); break;
+ case 'C': vc.min_lrt = atof(optarg); break;
+ case 'X': vc.min_perm_p = atof(optarg); break;
+ case 'd': vc.min_smpl_frac = atof(optarg); break;
+ case 's': vc.subsam = read_samples(optarg, &vc.n_sub);
+ vc.ploidy = calloc(vc.n_sub + 1, 1);
+ for (tid = 0; tid < vc.n_sub; ++tid) vc.ploidy[tid] = vc.subsam[tid][strlen(vc.subsam[tid]) + 1];
+ tid = -1;
+ break;
+ case 'T':
+ if (strcmp(optarg, "trioauto") == 0) vc.trio_aux = bcf_trio_prep(0, 0);
+ else if (strcmp(optarg, "trioxd") == 0) vc.trio_aux = bcf_trio_prep(1, 0);
+ else if (strcmp(optarg, "trioxs") == 0) vc.trio_aux = bcf_trio_prep(1, 1);
+ else if (strcmp(optarg, "pair") == 0) vc.flag |= VC_PAIRCALL;
+ else {
+ fprintf(stderr, "[%s] Option '-T' can only take value trioauto, trioxd or trioxs.\n", __func__);
+ return 1;
+ }
+ break;
case 'P':
if (strcmp(optarg, "full") == 0) vc.prior_type = MC_PTYPE_FULL;
else if (strcmp(optarg, "cond2") == 0) vc.prior_type = MC_PTYPE_COND2;
}
if (argc == optind) {
fprintf(stderr, "\n");
- fprintf(stderr, "Usage: bcftools view [options] <in.bcf> [reg]\n\n");
- fprintf(stderr, "Options: -c SNP calling\n");
- fprintf(stderr, " -v output potential variant sites only (force -c)\n");
- fprintf(stderr, " -g call genotypes at variant sites (force -c)\n");
- fprintf(stderr, " -b output BCF instead of VCF\n");
- fprintf(stderr, " -u uncompressed BCF output (force -b)\n");
- fprintf(stderr, " -S input is VCF\n");
- fprintf(stderr, " -A keep all possible alternate alleles at variant sites\n");
- fprintf(stderr, " -G suppress all individual genotype information\n");
- fprintf(stderr, " -H perform Hardy-Weinberg test (slower)\n");
- fprintf(stderr, " -N skip sites where REF is not A/C/G/T\n");
- fprintf(stderr, " -Q output the QCALL likelihood format\n");
- fprintf(stderr, " -L calculate LD for adjacent sites\n");
- fprintf(stderr, " -I skip indels\n");
- fprintf(stderr, " -1 INT number of group-1 samples [0]\n");
- fprintf(stderr, " -l FILE list of sites to output [all sites]\n");
- fprintf(stderr, " -t FLOAT scaled substitution mutation rate [%.4lg]\n", vc.theta);
- fprintf(stderr, " -i FLOAT indel-to-substitution ratio [%.4lg]\n", vc.indel_frac);
- fprintf(stderr, " -p FLOAT variant if P(ref|D)<FLOAT [%.3lg]\n", vc.pref);
- fprintf(stderr, " -P STR type of prior: full, cond2, flat [full]\n");
+ fprintf(stderr, "Usage: bcftools view [options] <in.bcf> [reg]\n\n");
+ fprintf(stderr, "Input/output options:\n\n");
+ fprintf(stderr, " -A keep all possible alternate alleles at variant sites\n");
+ fprintf(stderr, " -b output BCF instead of VCF\n");
+ fprintf(stderr, " -D FILE sequence dictionary for VCF->BCF conversion [null]\n");
+ fprintf(stderr, " -F PL generated by r921 or before (which generate old ordering)\n");
+ fprintf(stderr, " -G suppress all individual genotype information\n");
+ fprintf(stderr, " -l FILE list of sites (chr pos) or regions (BED) to output [all sites]\n");
+ fprintf(stderr, " -L calculate LD for adjacent sites\n");
+ fprintf(stderr, " -N skip sites where REF is not A/C/G/T\n");
+ fprintf(stderr, " -Q output the QCALL likelihood format\n");
+ fprintf(stderr, " -s FILE list of samples to use [all samples]\n");
+ fprintf(stderr, " -S input is VCF\n");
+ fprintf(stderr, " -u uncompressed BCF output (force -b)\n");
+ fprintf(stderr, "\nConsensus/variant calling options:\n\n");
+ fprintf(stderr, " -c SNP calling (force -e)\n");
+ fprintf(stderr, " -d FLOAT skip loci where less than FLOAT fraction of samples covered [0]\n");
+ fprintf(stderr, " -e likelihood based analyses\n");
+ fprintf(stderr, " -g call genotypes at variant sites (force -c)\n");
+ fprintf(stderr, " -i FLOAT indel-to-substitution ratio [%.4g]\n", vc.indel_frac);
+ fprintf(stderr, " -I skip indels\n");
+ fprintf(stderr, " -p FLOAT variant if P(ref|D)<FLOAT [%.3g]\n", vc.pref);
+ fprintf(stderr, " -P STR type of prior: full, cond2, flat [full]\n");
+ fprintf(stderr, " -t FLOAT scaled substitution mutation rate [%.4g]\n", vc.theta);
+ fprintf(stderr, " -T STR constrained calling; STR can be: pair, trioauto, trioxd and trioxs (see manual) [null]\n");
+ fprintf(stderr, " -v output potential variant sites only (force -c)\n");
+ fprintf(stderr, "\nContrast calling and association test options:\n\n");
+ fprintf(stderr, " -1 INT number of group-1 samples [0]\n");
+ fprintf(stderr, " -C FLOAT posterior constrast for LRT<FLOAT and P(ref|D)<0.5 [%g]\n", vc.min_lrt);
+ fprintf(stderr, " -U INT number of permutations for association testing (effective with -1) [0]\n");
+ fprintf(stderr, " -X FLOAT only perform permutations for P(chi^2)<FLOAT [%g]\n", vc.min_perm_p);
fprintf(stderr, "\n");
return 1;
}
+ if (vc.flag & VC_CALL) vc.flag |= VC_EM;
+ if ((vc.flag & VC_VCFIN) && (vc.flag & VC_BCFOUT) && vc.fn_dict == 0) {
+ fprintf(stderr, "[%s] For VCF->BCF conversion please specify the sequence dictionary with -D\n", __func__);
+ return 1;
+ }
+ if (vc.n1 <= 0) vc.n_perm = 0; // TODO: give a warning here!
+ if (vc.n_perm > 0) {
+ seeds = malloc(vc.n_perm * sizeof(int));
+ srand48(time(0));
+ for (c = 0; c < vc.n_perm; ++c) seeds[c] = lrand48();
+ }
b = calloc(1, sizeof(bcf1_t));
blast = calloc(1, sizeof(bcf1_t));
strcpy(moder, "r");
if (vc.flag & VC_BCFOUT) strcat(modew, "b");
if (vc.flag & VC_UNCOMP) strcat(modew, "u");
bp = vcf_open(argv[optind], moder);
- h = vcf_hdr_read(bp);
+ hin = hout = vcf_hdr_read(bp);
+ if (vc.fn_dict && (vc.flag & VC_VCFIN))
+ vcf_dictread(bp, hin, vc.fn_dict);
bout = vcf_open("-", modew);
- if (!(vc.flag & VC_QCALL)) vcf_hdr_write(bout, h);
+ if (!(vc.flag & VC_QCALL)) {
+ if (vc.n_sub) {
+ vc.sublist = calloc(vc.n_sub, sizeof(int));
+ hout = bcf_hdr_subsam(hin, vc.n_sub, vc.subsam, vc.sublist);
+ }
+ if (vc.flag & VC_CALL) write_header(hout);
+ vcf_hdr_write(bout, hout);
+ }
if (vc.flag & VC_CALL) {
- p1 = bcf_p1_init(h->n_smpl);
+ p1 = bcf_p1_init(hout->n_smpl, vc.ploidy);
if (vc.prior_file) {
if (bcf_p1_read_prior(p1, vc.prior_file) < 0) {
fprintf(stderr, "[%s] fail to read the prior AFS.\n", __func__);
return 1;
}
} else bcf_p1_init_prior(p1, vc.prior_type, vc.theta);
- if (vc.n1 > 0) {
+ if (vc.n1 > 0 && vc.min_lrt > 0.) { // set n1
bcf_p1_set_n1(p1, vc.n1);
bcf_p1_init_subprior(p1, vc.prior_type, vc.theta);
}
- if (vc.indel_frac > 0.) bcf_p1_indel_prior(p1, vc.indel_frac);
+ if (vc.indel_frac > 0.) bcf_p1_indel_prior(p1, vc.indel_frac); // otherwise use the default indel_frac
}
- if (vc.fn_list) hash = bcf_load_pos(vc.fn_list, h);
if (optind + 1 < argc && !(vc.flag&VC_VCFIN)) {
- void *str2id = bcf_build_refhash(h);
+ void *str2id = bcf_build_refhash(hout);
if (bcf_parse_region(str2id, argv[optind+1], &tid, &begin, &end) >= 0) {
bcf_idx_t *idx;
idx = bcf_idx_load(argv[optind]);
}
}
}
- while (vcf_read(bp, h, b) > 0) {
- int is_indel = bcf_is_indel(b);
+ while (vcf_read(bp, hin, b) > 0) {
+ int is_indel, cons_llr = -1;
+ int64_t cons_gt = -1;
+ double em[10];
+ if ((vc.flag & VC_VARONLY) && strcmp(b->alt, "X") == 0) continue;
+ if ((vc.flag & VC_VARONLY) && vc.min_smpl_frac > 0.) {
+ extern int bcf_smpl_covered(const bcf1_t *b);
+ int n = bcf_smpl_covered(b);
+ if ((double)n / b->n_smpl < vc.min_smpl_frac) continue;
+ }
+ if (vc.n_sub) bcf_subsam(vc.n_sub, vc.sublist, b);
+ if (vc.flag & VC_FIX_PL) bcf_fix_pl(b);
+ is_indel = bcf_is_indel(b);
if ((vc.flag & VC_NO_INDEL) && is_indel) continue;
if ((vc.flag & VC_ACGT_ONLY) && !is_indel) {
int x;
x = toupper(b->ref[0]);
if (x != 'A' && x != 'C' && x != 'G' && x != 'T') continue;
}
- if (hash) {
- uint64_t x = (uint64_t)b->tid<<32 | b->pos;
- khint_t k = kh_get(set64, hash, x);
- if (kh_size(hash) == 0) break;
- if (k == kh_end(hash)) continue;
- kh_del(set64, hash, k);
- }
+ if (vc.bed && !bed_overlap(vc.bed, hin->ns[b->tid], b->pos, b->pos + strlen(b->ref))) continue;
if (tid >= 0) {
int l = strlen(b->ref);
l = b->pos + (l > 0? l : 1);
if (!(l > begin && end > b->pos)) continue;
}
++n_processed;
+ if ((vc.flag & VC_QCNT) && !is_indel) { // summarize the difference
+ int x = bcf_min_diff(b);
+ if (x > 255) x = 255;
+ if (x >= 0) ++qcnt[x];
+ }
if (vc.flag & VC_QCALL) { // output QCALL format; STOP here
- bcf_2qcall(h, b);
+ bcf_2qcall(hout, b);
continue;
}
- if (vc.flag & (VC_CALL|VC_ADJLD)) bcf_gl2pl(b);
+ if (vc.trio_aux) // do trio calling
+ bcf_trio_call(vc.trio_aux, b, &cons_llr, &cons_gt);
+ else if (vc.flag & VC_PAIRCALL)
+ cons_llr = bcf_pair_call(b);
+ if (vc.flag & (VC_CALL|VC_ADJLD|VC_EM)) bcf_gl2pl(b);
+ if (vc.flag & VC_EM) bcf_em1(b, vc.n1, 0x1ff, em);
+ else {
+ int i;
+ for (i = 0; i < 9; ++i) em[i] = -1.;
+ }
if (vc.flag & VC_CALL) { // call variants
bcf_p1rst_t pr;
- bcf_p1_cal(b, p1, &pr); // pr.g[3] is not calculated here
- if (vc.flag&VC_HWE) bcf_p1_cal_g3(p1, pr.g);
+ int calret = bcf_p1_cal(b, (em[7] >= 0 && em[7] < vc.min_lrt), p1, &pr);
if (n_processed % 100000 == 0) {
fprintf(stderr, "[%s] %ld sites processed.\n", __func__, (long)n_processed);
bcf_p1_dump_afs(p1);
}
if (pr.p_ref >= vc.pref && (vc.flag & VC_VARONLY)) continue;
- update_bcf1(h->n_smpl, b, p1, &pr, vc.pref, vc.flag);
- }
+ if (vc.n_perm && vc.n1 > 0 && pr.p_chi2 < vc.min_perm_p) { // permutation test
+ bcf_p1rst_t r;
+ int i, n = 0;
+ for (i = 0; i < vc.n_perm; ++i) {
+#ifdef BCF_PERM_LRT // LRT based permutation is much faster but less robust to artifacts
+ double x[10];
+ bcf_shuffle(b, seeds[i]);
+ bcf_em1(b, vc.n1, 1<<7, x);
+ if (x[7] < em[7]) ++n;
+#else
+ bcf_shuffle(b, seeds[i]);
+ bcf_p1_cal(b, 1, p1, &r);
+ if (pr.p_chi2 >= r.p_chi2) ++n;
+#endif
+ }
+ pr.perm_rank = n;
+ }
+ if (calret >= 0) update_bcf1(b, p1, &pr, vc.pref, vc.flag, em, cons_llr, cons_gt);
+ } else if (vc.flag & VC_EM) update_bcf1(b, 0, 0, 0, vc.flag, em, cons_llr, cons_gt);
if (vc.flag & VC_ADJLD) { // compute LD
double f[4], r2;
- if ((r2 = bcf_ld_freq(blast, b, f)) >= 0) {
+ if ((r2 = bcf_pair_freq(blast, b, f)) >= 0) {
kstring_t s;
s.m = s.l = 0; s.s = 0;
if (*b->info) kputc(';', &s);
- ksprintf(&s, "NEIR=%.3lf;NEIF=%.3lf,%.3lf", r2, f[0]+f[2], f[0]+f[1]);
+ ksprintf(&s, "NEIR=%.3f;NEIF4=%.3f,%.3f,%.3f,%.3f", r2, f[0], f[1], f[2], f[3]);
bcf_append_info(b, s.s, s.l);
free(s.s);
}
bcf_cpy(blast, b);
}
+ if (vc.flag & VC_ANNO_MAX) bcf_anno_max(b);
if (vc.flag & VC_NO_GENO) { // do not output GENO fields
b->n_gi = 0;
b->fmt[0] = '\0';
- }
- vcf_write(bout, h, b);
+ b->l_str = b->fmt - b->str + 1;
+ } else bcf_fix_gt(b);
+ vcf_write(bout, hout, b);
}
if (vc.prior_file) free(vc.prior_file);
if (vc.flag & VC_CALL) bcf_p1_dump_afs(p1);
- bcf_hdr_destroy(h);
+ if (hin != hout) bcf_hdr_destroy(hout);
+ bcf_hdr_destroy(hin);
bcf_destroy(b); bcf_destroy(blast);
vcf_close(bp); vcf_close(bout);
- if (hash) kh_destroy(set64, hash);
- if (vc.fn_list) free(vc.fn_list);
+ if (vc.fn_dict) free(vc.fn_dict);
+ if (vc.ploidy) free(vc.ploidy);
+ if (vc.trio_aux) free(vc.trio_aux);
+ if (vc.n_sub) {
+ int i;
+ for (i = 0; i < vc.n_sub; ++i) free(vc.subsam[i]);
+ free(vc.subsam); free(vc.sublist);
+ }
+ if (vc.bed) bed_destroy(vc.bed);
+ if (vc.flag & VC_QCNT)
+ for (c = 0; c < 256; ++c)
+ fprintf(stderr, "QT\t%d\t%lld\n", c, (long long)qcnt[c]);
+ if (seeds) free(seeds);
if (p1) bcf_p1_destroy(p1);
return 0;
}
projects / samtools.git / blobdiff