#include #include #include #include #include "bntseq.h" #include "bwt.h" #include "kvec.h" #include "kseq.h" KSEQ_INIT(gzFile, gzread) typedef struct { int qbeg, len; int64_t tbeg; } fmhit_t; #define hit_qlt(a, b) ((a).qbeg < (b).qbeg) #define hit_tlt(a, b) ((a).tbeg < (b).tbeg) #include "ksort.h" KSORT_INIT(hitq, fmhit_t, hit_qlt) KSORT_INIT(hitt, fmhit_t, hit_tlt) extern unsigned char nst_nt4_table[256]; typedef struct { size_t n, m; fmhit_t *a; } fmhit_v; static uint64_t cluster_qhits(fmhit_v *hits, int beg, int end, int max_dist, int *_score) { int qend, score, max = 0, cbeg = beg; size_t i; uint64_t max_cluster = 0; ks_introsort(hitq, end - beg, hits->a + beg); qend = hits->a[beg].qbeg + hits->a[beg].len; score = hits->a[beg].len; for (i = beg + 1; i < end; ++i) { fmhit_t *p = &hits->a[i]; if (p->qbeg - qend > max_dist) { if (score > max) max = score, max_cluster = (uint64_t)cbeg<<32 | i; score = 0; cbeg = i; } score += p->len; qend = p->qbeg + p->len; } if (score > max) max = score, max_cluster = (uint64_t)cbeg<<32 | i; *_score = score; return max_cluster; } static int cluster_hits(fmhit_v *hits, int max_dist) { size_t i; int64_t tend; uint64_t cluster, max_cluster = 0; int j, score, max = 0, max2 = 0, cbeg = 0, cend; ks_introsort(hitt, hits->n, hits->a); tend = hits->a[0].tbeg + hits->a[0].len; for (i = 1; i < hits->n; ++i) { fmhit_t *p = &hits->a[i]; if (p->tbeg - tend > max_dist) { cluster = cluster_qhits(hits, cbeg, i, max_dist, &score); if (score > max) max2 = max, max = score, max_cluster = cluster; else if (score > max2) max2 = score; cbeg = i; } tend = p->tbeg + p->len; } cluster = cluster_qhits(hits, cbeg, i, max_dist, &score); if (score > max) max2 = max, max = score, max_cluster = cluster; else if (score > max2) max2 = score; cbeg = max_cluster>>32; cend = (uint32_t)max_cluster; for (i = 0, j = cbeg; j < cend; ++j) hits->a[i++] = hits->a[j]; hits->n = i; return (int)(200.0 * (max - max2) / max + .499); } static int fake_cigar(const bntseq_t *bns, fmhit_v *hits, int beg, int end, int len, uint32_t *cigar, int64_t *pos, int *is_rev) { size_t i; int qbeg, qend, n_cigar = 0; int64_t tbeg, tend, tmp; qbeg = len; qend = 0; tbeg = bns->l_pac<<1; tend = 0; for (i = beg; i < end; ++i) { fmhit_t *p = &hits->a[i]; if (p->qbeg < qbeg) qbeg = p->qbeg; if (p->qbeg + p->len > qend) qend = p->qbeg + p->len; if (p->tbeg < tbeg) tbeg = p->tbeg; if (p->tbeg + p->len > qend) tend = p->tbeg + p->len; } if (tbeg >= bns->l_pac) { tmp = tend; tend = bns->l_pac*2 - tbeg; tbeg = bns->l_pac*2 - tmp; tmp = qend; qend = len - qbeg; qbeg = len - tmp; *is_rev = 1; } else *is_rev = 0; *pos = tbeg; if (qbeg) cigar[n_cigar++] = qbeg<<4|4; if (tend - tbeg < qend - qbeg) { // reference is shorter cigar[n_cigar++] = (uint32_t)(tend - tbeg)<<4 | 0; cigar[n_cigar++] = (uint32_t)((qend - qbeg) - (tend - tbeg))<<4 | 2; } else if (tend - tbeg > qend - qbeg) { // query is shorter cigar[n_cigar++] = (uint32_t)(qend - qbeg)<<4 | 0; cigar[n_cigar++] = (uint32_t)((tend - tbeg) - (qend - qbeg))<<4 | 1; } else cigar[n_cigar++] = (uint32_t)(qend - qbeg)<<4 | 0; if (len > qend) cigar[n_cigar++] = (uint32_t)(len - qend)<<4|4; return n_cigar; } int main_fastmap(int argc, char *argv[]) { int c, i, min_iwidth = 20, min_len = 17, max_dist = 50, mem_only = 0; kseq_t *seq; bwtint_t k; gzFile fp; bwt_t *bwt; bntseq_t *bns; bwtintv_v a[3], mem, *tvec[3]; fmhit_v hits; uint32_t cigar[1024]; while ((c = getopt(argc, argv, "w:l:d:p")) >= 0) { switch (c) { case 'w': min_iwidth = atoi(optarg); break; case 'l': min_len = atoi(optarg); break; case 'd': max_dist = atoi(optarg); break; case 'p': mem_only = 1; break; } } if (optind + 1 >= argc) { fprintf(stderr, "bwa fastmap \n"); return 1; } fp = gzopen(argv[optind + 1], "r"); seq = kseq_init(fp); { // load the packed sequences, BWT and SA char *tmp = calloc(strlen(argv[optind]) + 5, 1); strcat(strcpy(tmp, argv[optind]), ".bwt"); bwt = bwt_restore_bwt(tmp); strcat(strcpy(tmp, argv[optind]), ".sa"); bwt_restore_sa(tmp, bwt); free(tmp); bns = bns_restore(argv[optind]); } for (i = 0; i < 3; ++i) { // initiate the temporary array kv_init(a[i]); tvec[i] = &a[i]; } kv_init(mem); kv_init(hits); while (kseq_read(seq) >= 0) { for (i = 0; i < seq->seq.l; ++i) seq->seq.s[i] = nst_nt4_table[(int)seq->seq.s[i]]; bwt_smem(bwt, seq->seq.l, (uint8_t*)seq->seq.s, &mem, tvec); if (mem_only) printf("SQ\t%s\t%ld\n", seq->name.s, seq->seq.l); for (i = 0, hits.n = 0; i < mem.n; ++i) { bwtintv_t *p = &mem.a[i]; if ((uint32_t)p->info - (p->info>>32) < min_len) continue; if (mem_only) printf("EM\t%d\t%d\t%ld", (uint32_t)(p->info>>32), (uint32_t)p->info, (long)p->x[2]); if (p->x[2] <= min_iwidth) { for (k = 0; k < p->x[2]; ++k) { fmhit_t z; z.tbeg = bwt_sa(bwt, p->x[0] + k); z.qbeg = p->info>>32; z.len = (uint32_t)p->info - z.qbeg; kv_push(fmhit_t, hits, z); if (mem_only) { int is_rev, ref_id; int64_t pos = bns_depos(bns, z.tbeg, &is_rev); if (is_rev) pos -= z.len - 1; bns_cnt_ambi(bns, pos, z.len, &ref_id); printf("\t%s:%c%ld", bns->anns[ref_id].name, "+-"[is_rev], (long)(pos - bns->anns[ref_id].offset) + 1); } } } if (mem_only) putchar('\n'); } if (!mem_only) { int64_t pos; int n_cigar, is_rev, ref_id, mapq; if (hits.n == 0) continue; mapq = cluster_hits(&hits, max_dist); n_cigar = fake_cigar(bns, &hits, 0, hits.n, seq->seq.l, cigar, &pos, &is_rev); bns_cnt_ambi(bns, pos, 1, &ref_id); printf("%s\t%d\t%s\t%ld\t%d\t", seq->name.s, is_rev?16:0, bns->anns[ref_id].name, (long)(pos - bns->anns[ref_id].offset) + 1, mapq); for (i = 0; i < n_cigar; ++i) printf("%d%c", cigar[i]>>4, "MIDNSHP"[cigar[i]&0xf]); printf("\t*\t0\t0\t*\t*\n"); } else puts("//"); } free(mem.a); for (i = 0; i < 3; ++i) free(a[i].a); bns_destroy(bns); bwt_destroy(bwt); kseq_destroy(seq); gzclose(fp); return 0; }