/* bam_tview.c -- tview subcommand. Copyright (C) 2008-2014 Genome Research Ltd. Portions copyright (C) 2013 Pierre Lindenbaum, Institut du Thorax, INSERM U1087, Université de Nantes. Author: Heng Li Permission is hereby granted, free of charge, to any person obtaining a copy of this software and associated documentation files (the "Software"), to deal in the Software without restriction, including without limitation the rights to use, copy, modify, merge, publish, distribute, sublicense, and/or sell copies of the Software, and to permit persons to whom the Software is furnished to do so, subject to the following conditions: The above copyright notices and this permission notice shall be included in all copies or substantial portions of the Software. THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM, OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE SOFTWARE. */ #include #include #include "bam_tview.h" #include #include #include /*! @typedef @abstract Type of function to be called by sam_fetch(). @param b the alignment @param data user provided data */ typedef int (*sam_fetch_f)(const bam1_t *b, void *data); int sam_fetch(samFile *fp, const hts_idx_t *idx, int tid, int beg, int end, void *data, sam_fetch_f func) { int ret; hts_itr_t* iter; bam1_t* b = bam_init1(); iter = sam_itr_queryi(idx, tid, beg, end); while ((ret = sam_itr_next(fp, iter, b)) >= 0) func(b, data); hts_itr_destroy(iter); bam_destroy1(b); return (ret == -1)? 0 : ret; } khash_t(kh_rg)* get_rg_sample(const char* header, const char* sample) { khash_t(kh_rg)* rg_hash = kh_init(kh_rg); // given sample id return all the RD ID's const char rg_regex[] = "^@RG.*\tID:([!-)+-<>-~][ !-~]*)(\t.*$|$)"; regex_t rg_id; regmatch_t* matches = (regmatch_t*)calloc(2, sizeof(regmatch_t)); if (matches == NULL) { perror("out of memory"); exit(-1); } regcomp(&rg_id, rg_regex, REG_EXTENDED|REG_NEWLINE); char* text = strdup(header); char* end = text + strlen(header); char* tofree = text; while (end > text && regexec(&rg_id, text, 2, matches, 0) == 0) { // foreach rg id in header int ret; text[matches[1].rm_eo] = '\0'; kh_put(kh_rg, rg_hash, strdup(text+matches[1].rm_so), &ret); // Add the RG to the list text += matches[0].rm_eo + 1; // Move search pointer forward } free(tofree); return rg_hash; } int base_tv_init(tview_t* tv, const char *fn, const char *fn_fa, const char *samples) { assert(tv!=NULL); assert(fn!=NULL); tv->mrow = 24; tv->mcol = 80; tv->color_for = TV_COLOR_MAPQ; tv->is_dot = 1; tv->fp = sam_open(fn, "r"); if(tv->fp == NULL) { fprintf(stderr,"sam_open %s. %s\n", fn,fn_fa); exit(EXIT_FAILURE); } // TODO bgzf_set_cache_size(tv->fp->fp.bgzf, 8 * 1024 *1024); assert(tv->fp); tv->header = sam_hdr_read(tv->fp); if(tv->header == NULL) { fprintf(stderr,"Cannot read '%s'.\n", fn); exit(EXIT_FAILURE); } tv->idx = sam_index_load(tv->fp, fn); if (tv->idx == NULL) { fprintf(stderr,"Cannot read index for '%s'.\n", fn); exit(EXIT_FAILURE); } tv->lplbuf = bam_lplbuf_init(tv_pl_func, tv); if (fn_fa) tv->fai = fai_load(fn_fa); tv->bca = bcf_call_init(0.83, 13); tv->ins = 1; // If the user has asked for specific samples find out create a list of readgroups make up these samples if ( samples ) { tv->rg_hash = get_rg_sample(tv->header->text, samples); // Init the list of rg's } return 0; } void base_tv_destroy(tview_t* tv) { bam_lplbuf_destroy(tv->lplbuf); bcf_call_destroy(tv->bca); hts_idx_destroy(tv->idx); if (tv->fai) fai_destroy(tv->fai); free(tv->ref); bam_hdr_destroy(tv->header); sam_close(tv->fp); } int tv_pl_func(uint32_t tid, uint32_t pos, int n, const bam_pileup1_t *pl, void *data) { extern const char bam_nt16_nt4_table[]; tview_t *tv = (tview_t*)data; int i, j, c, rb, attr, max_ins = 0; uint32_t call = 0; if (pos < tv->left_pos || tv->ccol > tv->mcol) return 0; // out of screen // print reference rb = (tv->ref && pos - tv->left_pos < tv->l_ref)? tv->ref[pos - tv->left_pos] : 'N'; for (i = tv->last_pos + 1; i < pos; ++i) { if (i%10 == 0 && tv->mcol - tv->ccol >= 10) tv->my_mvprintw(tv,0, tv->ccol, "%-d", i+1); c = tv->ref? tv->ref[i - tv->left_pos] : 'N'; tv->my_mvaddch(tv,1, tv->ccol++, c); } if (pos%10 == 0 && tv->mcol - tv->ccol >= 10) tv->my_mvprintw(tv,0, tv->ccol, "%-d", pos+1); { // call consensus bcf_callret1_t bcr; memset(&bcr, 0, sizeof bcr); int qsum[4], a1, a2, tmp; double p[3], prior = 30; bcf_call_glfgen(n, pl, seq_nt16_table[rb], tv->bca, &bcr); for (i = 0; i < 4; ++i) qsum[i] = ((int)bcr.qsum[i])<<2 | i; for (i = 1; i < 4; ++i) // insertion sort for (j = i; j > 0 && qsum[j] > qsum[j-1]; --j) tmp = qsum[j], qsum[j] = qsum[j-1], qsum[j-1] = tmp; a1 = qsum[0]&3; a2 = qsum[1]&3; p[0] = bcr.p[a1*5+a1]; p[1] = bcr.p[a1*5+a2] + prior; p[2] = bcr.p[a2*5+a2]; if ("ACGT"[a1] != toupper(rb)) p[0] += prior + 3; if ("ACGT"[a2] != toupper(rb)) p[2] += prior + 3; if (p[0] < p[1] && p[0] < p[2]) call = (1<my_underline(tv); c = ",ACMGRSVTWYHKDBN"[call>>16&0xf]; i = (call&0xffff)/10+1; if (i > 4) i = 4; attr |= tv->my_colorpair(tv,i); if (c == toupper(rb)) c = '.'; tv->my_attron(tv,attr); tv->my_mvaddch(tv,2, tv->ccol, c); tv->my_attroff(tv,attr); if(tv->ins) { // calculate maximum insert for (i = 0; i < n; ++i) { const bam_pileup1_t *p = pl + i; if (p->indel > 0 && max_ins < p->indel) max_ins = p->indel; } } // core loop for (j = 0; j <= max_ins; ++j) { for (i = 0; i < n; ++i) { const bam_pileup1_t *p = pl + i; int row = TV_MIN_ALNROW + p->level - tv->row_shift; if (j == 0) { if (!p->is_del) { if (tv->base_for == TV_BASE_COLOR_SPACE && (c = bam_aux_getCSi(p->b, p->qpos))) { // assume that if we found one color, we will be able to get the color error if (tv->is_dot && '-' == bam_aux_getCEi(p->b, p->qpos)) c = bam_is_rev(p->b)? ',' : '.'; } else { if (tv->show_name) { char *name = bam_get_qname(p->b); c = (p->qpos + 1 >= p->b->core.l_qname)? ' ' : name[p->qpos]; } else { c = seq_nt16_str[bam_seqi(bam_get_seq(p->b), p->qpos)]; if (tv->is_dot && toupper(c) == toupper(rb)) c = bam_is_rev(p->b)? ',' : '.'; } } } else c = p->is_refskip? (bam_is_rev(p->b)? '<' : '>') : '*'; } else { // padding if (j > p->indel) c = '*'; else { // insertion if (tv->base_for == TV_BASE_NUCL) { if (tv->show_name) { char *name = bam_get_qname(p->b); c = (p->qpos + j + 1 >= p->b->core.l_qname)? ' ' : name[p->qpos + j]; } else { c = seq_nt16_str[bam_seqi(bam_get_seq(p->b), p->qpos + j)]; if (j == 0 && tv->is_dot && toupper(c) == toupper(rb)) c = bam_is_rev(p->b)? ',' : '.'; } } else { c = bam_aux_getCSi(p->b, p->qpos + j); if (tv->is_dot && '-' == bam_aux_getCEi(p->b, p->qpos + j)) c = bam_is_rev(p->b)? ',' : '.'; } } } if (row > TV_MIN_ALNROW && row < tv->mrow) { int x; attr = 0; if (((p->b->core.flag&BAM_FPAIRED) && !(p->b->core.flag&BAM_FPROPER_PAIR)) || (p->b->core.flag & BAM_FSECONDARY)) attr |= tv->my_underline(tv); if (tv->color_for == TV_COLOR_BASEQ) { x = bam_get_qual(p->b)[p->qpos]/10 + 1; if (x > 4) x = 4; attr |= tv->my_colorpair(tv,x); } else if (tv->color_for == TV_COLOR_MAPQ) { x = p->b->core.qual/10 + 1; if (x > 4) x = 4; attr |= tv->my_colorpair(tv,x); } else if (tv->color_for == TV_COLOR_NUCL) { x = bam_nt16_nt4_table[bam_seqi(bam_get_seq(p->b), p->qpos)] + 5; attr |= tv->my_colorpair(tv,x); } else if(tv->color_for == TV_COLOR_COL) { x = 0; switch(bam_aux_getCSi(p->b, p->qpos)) { case '0': x = 0; break; case '1': x = 1; break; case '2': x = 2; break; case '3': x = 3; break; case '4': x = 4; break; default: x = bam_nt16_nt4_table[bam_seqi(bam_get_seq(p->b), p->qpos)]; break; } x+=5; attr |= tv->my_colorpair(tv,x); } else if(tv->color_for == TV_COLOR_COLQ) { x = bam_aux_getCQi(p->b, p->qpos); if(0 == x) x = bam_get_qual(p->b)[p->qpos]; x = x/10 + 1; if (x > 4) x = 4; attr |= tv->my_colorpair(tv,x); } tv->my_attron(tv,attr); tv->my_mvaddch(tv,row, tv->ccol, bam_is_rev(p->b)? tolower(c) : toupper(c)); tv->my_attroff(tv,attr); } } c = j? '*' : rb; if (c == '*') { attr = tv->my_colorpair(tv,8); tv->my_attron(tv,attr); tv->my_mvaddch(tv,1, tv->ccol++, c); tv->my_attroff(tv,attr); } else tv->my_mvaddch(tv,1, tv->ccol++, c); } tv->last_pos = pos; return 0; } int tv_fetch_func(const bam1_t *b, void *data) { tview_t *tv = (tview_t*)data; /* If we are restricted to specific readgroups check RG is in the list */ if ( tv->rg_hash ) { const uint8_t *rg = bam_aux_get(b, "RG"); if ( !rg ) return 0; // If we don't have an RG tag exclude read khiter_t k = kh_get(kh_rg, tv->rg_hash, (const char*)(rg + 1)); if ( k == kh_end(tv->rg_hash) ) return 0; // if RG tag is not in list of allowed tags exclude read } if (tv->no_skip) { uint32_t *cigar = bam_get_cigar(b); // this is cheating... int i; for (i = 0; i core.n_cigar; ++i) { if ((cigar[i]&0xf) == BAM_CREF_SKIP) cigar[i] = cigar[i]>>4<<4 | BAM_CDEL; } } bam_lplbuf_push(b, tv->lplbuf); return 0; } int base_draw_aln(tview_t *tv, int tid, int pos) { assert(tv!=NULL); // reset tv->my_clear(tv); tv->curr_tid = tid; tv->left_pos = pos; tv->last_pos = tv->left_pos - 1; tv->ccol = 0; // print ref and consensus if (tv->fai) { char *str; if (tv->ref) free(tv->ref); assert(tv->curr_tid>=0); str = (char*)calloc(strlen(tv->header->target_name[tv->curr_tid]) + 30, 1); assert(str!=NULL); sprintf(str, "%s:%d-%d", tv->header->target_name[tv->curr_tid], tv->left_pos + 1, tv->left_pos + tv->mcol); tv->ref = fai_fetch(tv->fai, str, &tv->l_ref); free(str); if ( !tv->ref ) { fprintf(stderr,"Could not read the reference sequence. Is it seekable (plain text or compressed + .gzi indexed with bgzip)?\n"); exit(1); } } // draw aln bam_lplbuf_reset(tv->lplbuf); sam_fetch(tv->fp, tv->idx, tv->curr_tid, tv->left_pos, tv->left_pos + tv->mcol, tv, tv_fetch_func); bam_lplbuf_push(0, tv->lplbuf); while (tv->ccol < tv->mcol) { int pos = tv->last_pos + 1; if (pos%10 == 0 && tv->mcol - tv->ccol >= 10) tv->my_mvprintw(tv,0, tv->ccol, "%-d", pos+1); tv->my_mvaddch(tv,1, tv->ccol++, (tv->ref && pos < tv->l_ref)? tv->ref[pos - tv->left_pos] : 'N'); ++tv->last_pos; } return 0; } static void error(const char *format, ...) { if ( !format ) { fprintf(stderr, "Usage: samtools tview [options] [ref.fasta]\n" "Options:\n" " -d display output as (H)tml or (C)urses or (T)ext \n" " -p chr:pos go directly to this position\n" " -s STR display only reads from this sample or group\n"); } else { va_list ap; va_start(ap, format); vfprintf(stderr, format, ap); va_end(ap); } exit(-1); } enum dipsay_mode {display_ncurses,display_html,display_text}; extern tview_t* curses_tv_init(const char *fn, const char *fn_fa, const char *samples); extern tview_t* html_tv_init(const char *fn, const char *fn_fa, const char *samples); extern tview_t* text_tv_init(const char *fn, const char *fn_fa, const char *samples); int bam_tview_main(int argc, char *argv[]) { int view_mode=display_ncurses; tview_t* tv=NULL; char *samples=NULL, *position=NULL; int c; while ((c = getopt(argc, argv, "s:p:d:")) >= 0) { switch (c) { case 's': samples=optarg; break; case 'p': position=optarg; break; case 'd': { switch(optarg[0]) { case 'H': case 'h': view_mode=display_html;break; case 'T': case 't': view_mode=display_text;break; case 'C': case 'c': view_mode=display_ncurses;break; default: view_mode=display_ncurses;break; } break; } default: error(NULL); } } if (argc==optind) error(NULL); switch(view_mode) { case display_ncurses: { tv = curses_tv_init(argv[optind], (optind+1>=argc)? 0 : argv[optind+1], samples); break; } case display_text: { tv = text_tv_init(argv[optind], (optind+1>=argc)? 0 : argv[optind+1], samples); break; } case display_html: { tv = html_tv_init(argv[optind], (optind+1>=argc)? 0 : argv[optind+1], samples); break; } } if (tv==NULL) { error("cannot create view"); return EXIT_FAILURE; } if ( position ) { int tid, beg, end; *(char *)hts_parse_reg(position, &beg, &end) = '\0'; tid = bam_name2id(tv->header, position); if (tid >= 0) { tv->curr_tid = tid; tv->left_pos = beg; } } else if ( tv->fai ) { // find the first sequence present in both BAM and the reference file int i; for (i=0; iheader->n_targets; i++) { if ( faidx_has_seq(tv->fai, tv->header->target_name[i]) ) break; } if ( i==tv->header->n_targets ) { fprintf(stderr,"None of the BAM sequence names present in the fasta file\n"); exit(EXIT_FAILURE); } tv->curr_tid = i; } tv->my_drawaln(tv, tv->curr_tid, tv->left_pos); tv->my_loop(tv); tv->my_destroy(tv); return EXIT_SUCCESS; }