#include "sam.h"
#include "faidx.h"
#include "bam_maqcns.h"
+#include "bam_mcns.h"
#include "khash.h"
#include "glf.h"
#include "kstring.h"
* mpileup *
***********/
+#define MPLP_VCF 0x1
+#define MPLP_VAR 0x2
+#define MPLP_AFS 0x4
+#define MPLP_AFALL 0x8
+
+#define MPLP_AFS_BLOCK 0x10000
+
typedef struct {
+ int max_mq, min_mq, prior_type, flag;
+ double theta;
char *reg, *fn_pos;
faidx_t *fai;
kh_64_t *hash;
typedef struct {
bamFile fp;
bam_iter_t iter;
+ int min_mq;
} mplp_aux_t;
static int mplp_func(void *data, bam1_t *b)
{
mplp_aux_t *ma = (mplp_aux_t*)data;
- if (ma->iter) return bam_iter_read(ma->fp, ma->iter, b);
- return bam_read1(ma->fp, b);
+ int ret;
+ do {
+ ret = ma->iter? bam_iter_read(ma->fp, ma->iter, b) : bam_read1(ma->fp, b);
+ } while (b->core.qual < ma->min_mq && ret >= 0);
+ return ret;
}
static int mpileup(mplp_conf_t *conf, int n, char **fn)
{
mplp_aux_t **data;
+ mc_aux_t *ma = 0;
int i, tid, pos, *n_plp, beg0 = 0, end0 = 1u<<29, ref_len, ref_tid;
const bam_pileup1_t **plp;
bam_mplp_t iter;
bam_header_t *h = 0;
+ uint64_t N = 0;
char *ref;
khash_t(64) *hash = 0;
// allocate
for (i = 0; i < n; ++i) {
bam_header_t *h_tmp;
data[i] = calloc(1, sizeof(mplp_aux_t));
+ data[i]->min_mq = conf->min_mq;
data[i]->fp = bam_open(fn[i], "r");
h_tmp = bam_header_read(data[i]->fp);
if (conf->reg) {
}
}
if (conf->fn_pos) hash = load_pos(conf->fn_pos, h);
+ // write the VCF header
+ if (conf->flag & MPLP_VCF) {
+ kstring_t s;
+ s.l = s.m = 0; s.s = 0;
+ puts("##fileformat=VCFv4.0");
+ puts("##INFO=<ID=DP,Number=1,Type=Integer,Description=\"Total read depth\">");
+ puts("##INFO=<ID=AF,Number=1,Type=Float,Description=\"Non-reference allele frequency \\argmax_f P(D|f)\">");
+ puts("##INFO=<ID=AFE,Number=1,Type=Float,Description=\"Expected non-reference allele frequency\">");
+ puts("##FILTER=<ID=Q13,Description=\"All min{baseQ,mapQ} below 13\">");
+ kputs("#CHROM\tPOS\tID\tREF\tALT\tQUAL\tFILTER\tINFO\tFORMAT", &s);
+ for (i = 0; i < n; ++i) {
+ const char *p;
+ kputc('\t', &s);
+ if ((p = strstr(fn[i], ".bam")) != 0)
+ kputsn(fn[i], p - fn[i], &s);
+ else kputs(fn[i], &s);
+ }
+ puts(s.s);
+ free(s.s);
+ }
// mpileup
+ if (conf->flag & MPLP_VCF) {
+ ma = mc_init(n);
+ mc_init_prior(ma, conf->prior_type, conf->theta);
+ }
ref_tid = -1; ref = 0;
iter = bam_mplp_init(n, mplp_func, (void**)data);
while (bam_mplp_auto(iter, &tid, &pos, n_plp, plp) > 0) {
if (conf->fai) ref = fai_fetch(conf->fai, h->target_name[tid], &ref_len);
ref_tid = tid;
}
- printf("%s\t%d\t%c", h->target_name[tid], pos + 1, (ref && pos < ref_len)? ref[pos] : 'N');
- for (i = 0; i < n; ++i) {
- int j;
- printf("\t%d\t", n_plp[i]);
- if (n_plp[i] == 0) printf("*\t*");
- else {
- for (j = 0; j < n_plp[i]; ++j)
- pileup_seq(plp[i] + j, pos, ref_len, ref);
- putchar('\t');
+ if (conf->flag & MPLP_VCF) {
+ mc_rst_t r;
+ int j, _ref0, depth, rms_q, _ref0b, is_var = 0, qref = 0, level = 2, tot;
+ uint64_t sqr_sum;
+ if (conf->flag & MPLP_AFS) level = 3;
+ _ref0 = _ref0b = (ref && pos < ref_len)? ref[pos] : 'N';
+ _ref0 = bam_nt16_nt4_table[bam_nt16_table[_ref0]];
+ tot = mc_cal(_ref0, n_plp, plp, ma, &r, level);
+ if (tot) { // has good bases
+ double q;
+ is_var = (r.p_ref < .5);
+ q = is_var? r.p_ref : 1. - r.p_ref;
+ if (q < 1e-308) q = 1e-308;
+ qref = (int)(-3.434 * log(q) + .499);
+ if (qref > 99) qref = 99;
+ }
+ if ((conf->flag & MPLP_VAR) && !is_var) continue;
+ ++N; // number of processed lines
+ printf("%s\t%d\t.\t%c\t", h->target_name[tid], pos + 1, _ref0b);
+ if (is_var) {
+ putchar("ACGTN"[r.alt]);
+ if (r.alt2 >= 0 && r.alt2 < 4) printf(",%c", "ACGT"[r.alt2]);
+ } else putchar('.');
+ printf("\t%d\t", qref);
+ if (!tot) printf("Q13\t");
+ else printf(".\t");
+ for (i = depth = 0, sqr_sum = 0; i < n; ++i) {
+ depth += n_plp[i];
for (j = 0; j < n_plp[i]; ++j) {
- const bam_pileup1_t *p = plp[i] + j;
- int c = bam1_qual(p->b)[p->qpos] + 33;
- if (c > 126) c = 126;
- putchar(c);
+ int q = plp[i][j].b->core.qual;
+ if (q > conf->max_mq) q = conf->max_mq;
+ sqr_sum += q * q;
+ }
+ }
+ rms_q = (int)(sqrt((double)sqr_sum / depth) + .499);
+ printf("DP=%d;MQ=%d", depth, rms_q);
+ if (tot) {
+ printf(";AF=%.3lf", 1. - r.f_em);
+ if (level >= 2) printf(";AFE=%.3lf", 1-r.f_exp);
+ if (conf->flag & MPLP_AFALL) {
+ printf(";AF0=%.3lf;AFN=%.3lf", 1-r.f_naive, 1-r.f_nielsen);
+ if (conf->flag & MPLP_AFS) printf(";AFB=%.3lf", 1-r.f_map);
+ }
+ }
+ printf("\tGT:GQ:DP");
+ if (tot) {
+ for (i = 0; i < n; ++i) {
+ int x = mc_call_gt(ma, r.f_exp, i);
+ printf("\t%c/%c:%d:%d", "10"[((x&3)==2)], "10"[((x&3)>0)], x>>2, n_plp[i]);
+ }
+ } else for (i = 0; i < n; ++i) printf("\t./.:0:0");
+ putchar('\n');
+ if (N % MPLP_AFS_BLOCK == 0) mc_dump_afs(ma);
+ } else {
+ printf("%s\t%d\t%c", h->target_name[tid], pos + 1, (ref && pos < ref_len)? ref[pos] : 'N');
+ for (i = 0; i < n; ++i) {
+ int j;
+ printf("\t%d\t", n_plp[i]);
+ if (n_plp[i] == 0) printf("*\t*");
+ else {
+ for (j = 0; j < n_plp[i]; ++j)
+ pileup_seq(plp[i] + j, pos, ref_len, ref);
+ putchar('\t');
+ for (j = 0; j < n_plp[i]; ++j) {
+ const bam_pileup1_t *p = plp[i] + j;
+ int c = bam1_qual(p->b)[p->qpos] + 33;
+ if (c > 126) c = 126;
+ putchar(c);
+ }
}
}
+ putchar('\n');
}
- putchar('\n');
}
+ if ((conf->flag&MPLP_VCF) && (conf->flag&MPLP_AFS)) mc_dump_afs(ma);
if (hash) { // free the hash table
khint_t k;
for (k = kh_begin(hash); k < kh_end(hash); ++k)
if (kh_exist(hash, k)) free(kh_val(hash, k));
kh_destroy(64, hash);
}
+ mc_destroy(ma);
bam_mplp_destroy(iter);
bam_header_destroy(h);
for (i = 0; i < n; ++i) {
int c;
mplp_conf_t mplp;
memset(&mplp, 0, sizeof(mplp_conf_t));
- while ((c = getopt(argc, argv, "f:r:l:")) >= 0) {
+ mplp.max_mq = 60;
+ mplp.prior_type = MC_PTYPE_FULL;
+ mplp.theta = 1e-3;
+ while ((c = getopt(argc, argv, "vVcFSP:f:r:l:VM:q:t:")) >= 0) {
switch (c) {
+ case 't': mplp.theta = atof(optarg); break;
+ case 'P':
+ if (strcmp(optarg, "full") == 0) mplp.prior_type = MC_PTYPE_FULL;
+ else if (strcmp(optarg, "cond2") == 0) mplp.prior_type = MC_PTYPE_COND2;
+ else if (strcmp(optarg, "flat") == 0) mplp.prior_type = MC_PTYPE_FLAT;
+ else {
+ fprintf(stderr, "[%s] unrecognized prior type.\n", __func__);
+ return 1;
+ }
+ break;
case 'f':
mplp.fai = fai_load(optarg);
if (mplp.fai == 0) return 1;
break;
case 'r': mplp.reg = strdup(optarg); break;
case 'l': mplp.fn_pos = strdup(optarg); break;
+ case 'V':
+ case 'c': mplp.flag |= MPLP_VCF; break;
+ case 'S': mplp.flag |= MPLP_AFS; break;
+ case 'F': mplp.flag |= MPLP_AFALL; break;
+ case 'v': mplp.flag |= MPLP_VAR; break;
+ case 'M': mplp.max_mq = atoi(optarg); break;
+ case 'q': mplp.min_mq = atoi(optarg); break;
}
}
if (argc == 1) {
- fprintf(stderr, "Usage: samtools mpileup [-r reg] [-f in.fa] [-l pos] in1.bam [in2.bam [...]]\n");
+ fprintf(stderr, "\n");
+ fprintf(stderr, "Usage: samtools mpileup [options] in1.bam [in2.bam [...]]\n\n");
+ fprintf(stderr, "Options: -f FILE reference sequence file [null]\n");
+ fprintf(stderr, " -r STR region in which pileup is generated [null]\n");
+ fprintf(stderr, " -l FILE list of positions (format: chr pos) [null]\n");
+ fprintf(stderr, " -M INT cap mapping quality at INT [%d]\n", mplp.max_mq);
+ fprintf(stderr, " -q INT filter out alignment with MQ smaller than INT [%d]\n", mplp.min_mq);
+ fprintf(stderr, " -t FLOAT scaled mutation rate [%lg]\n", mplp.theta);
+ fprintf(stderr, " -P STR prior: full, flat, cond2 [full]\n");
+ fprintf(stderr, " -c generate VCF output (consensus calling)\n");
+ fprintf(stderr, " -v show variant sites only\n");
+ fprintf(stderr, " -S calculate AFS (slow, to stderr)\n");
+ fprintf(stderr, "\n");
+ fprintf(stderr, "Notes: Assuming error independency and diploid individuals.\n\n");
return 1;
}
mpileup(&mplp, argc - optind, argv + optind);
projects / samtools.git / blobdiff