#include "sam.h"
#include "kstring.h"
#include "kaln.h"
+#include "kprobaln.h"
+
+char bam_nt16_nt4_table[] = { 4, 0, 1, 4, 2, 4, 4, 4, 3, 4, 4, 4, 4, 4, 4, 4 };
void bam_fillmd1_core(bam1_t *b, char *ref, int is_equal, int max_nm)
{
return (int)(t + .499);
}
-// local realignment
-
-#define MIN_REF_LEN 10
-#define MIN_BAND_WIDTH 11
-
-int bam_realn(bam1_t *b, const char *ref)
+int bam_prob_realn_core(bam1_t *b, const char *ref, int flag)
{
- int k, score, q[2], r[2], kk[2], kl[2], x, y, max, j, n_cigar, endx = -1;
+ int k, i, bw, x, y, yb, ye, xb, xe, apply_baq = flag&1, extend_baq = flag>>1&1;
uint32_t *cigar = bam1_cigar(b);
- uint8_t *seq = bam1_seq(b);
bam1_core_t *c = &b->core;
- q[0] = q[1] = r[0] = r[1] = kk[0] = kk[1] = kl[0] = kl[1] = -1;
- // find the right boundary
- for (k = 0, score = max = 0, x = c->pos, y = 0; k < c->n_cigar; ++k) {
- int op = cigar[k]&0xf;
- int ol = cigar[k]>>4;
- if (op == BAM_CMATCH) {
- for (j = 0; j < ol; ++j) {
- int c1, c2, z = y + j;
- c1 = bam_nt16_nt4_table[bam1_seqi(seq, z)];
- if (ref[x+j] == 0) return -1;
- c2 = bam_nt16_nt4_table[(int)bam_nt16_table[(int)ref[x+j]]];
- if (c1 < 3 && c2 < 3)
- score += c1 == c2? 5 : -4;
- if (score < 0) score = 0;
- if (score > max) max = score, q[1] = z, r[1] = x+j, kk[1] = k, kl[1] = j + 1;
- }
- x += ol; y += ol;
- } else if (op == BAM_CINS) {
- score -= 4 - ol * 3;
- y += ol;
- if (score < 0) score = 0;
- } else if (op == BAM_CDEL) {
- score -= 4 - ol * 3;
- x += ol;
- if (score < 0) score = 0;
- } else if (op == BAM_CSOFT_CLIP) y += ol;
- else if (op == BAM_CREF_SKIP) x += ol;
- }
- if (q[1] < 0) return -1; // no high scoring segments
- endx = x - 1;
- // find the left boundary
- for (k = c->n_cigar - 1, score = max = 0, x = x-1, y = y-1; k >= 0; --k) {
- int op = cigar[k]&0xf;
- int ol = cigar[k]>>4;
- if (op == BAM_CMATCH) {
- for (j = 0; j < ol; ++j) {
- int c1, c2, z = y - j;
- c1 = bam_nt16_nt4_table[bam1_seqi(seq, z)];
- if (ref[x+j] == 0) return -1;
- c2 = bam_nt16_nt4_table[(int)bam_nt16_table[(int)ref[x-j]]];
- if (c1 < 3 && c2 < 3)
- score += c1 == c2? 5 : -4;
- if (score < 0) score = 0;
- if (score > max) max = score, q[0] = z, r[0] = x-j, kk[0] = k, kl[0] = j + 1;
- }
- x -= ol; y -= ol;
- } else if (op == BAM_CINS) {
- score -= 4 - ol * 3;
- y -= ol;
- if (score < 0) score = 0;
- } else if (op == BAM_CDEL) {
- score -= 4 - ol * 3;
- x -= ol;
- if (score < 0) score = 0;
- } else if (op == BAM_CSOFT_CLIP) y -= ol;
- else if (op == BAM_CREF_SKIP) x -= ol;
+ kpa_par_t conf = kpa_par_def;
+ uint8_t *bq = 0, *zq = 0, *qual = bam1_qual(b);
+ if ((c->flag & BAM_FUNMAP) || b->core.l_qseq == 0) return -1; // do nothing
+ // test if BQ or ZQ is present
+ if ((bq = bam_aux_get(b, "BQ")) != 0) ++bq;
+ if ((zq = bam_aux_get(b, "ZQ")) != 0 && *zq == 'Z') ++zq;
+ if (bq && zq) { // remove the ZQ tag
+ bam_aux_del(b, zq-1);
+ zq = 0;
}
- if (q[0] < 0 || q[1] - q[0] < 15) return -1; // the high-scoring segment is too short
- // modify CIGAR
- n_cigar = 0;
- cigar = calloc(c->n_cigar + 4, 4);
- if (q[0] != 0) cigar[n_cigar++] = (uint32_t)q[0]<<4 | BAM_CSOFT_CLIP;
- if (r[0] != c->pos) cigar[n_cigar++] = (uint32_t)(r[0] - c->pos)<<4 | BAM_CREF_SKIP;
- if (kk[0] == kk[1]) {
- cigar[n_cigar++] = (uint32_t)(kl[0] + kl[1] - (bam1_cigar(b)[kk[0]]>>4))<<4 | BAM_CMATCH;
- } else {
- cigar[n_cigar++] = (uint32_t)kl[0]<<4 | BAM_CMATCH;
- for (k = kk[0] + 1; k < kk[1]; ++k)
- cigar[n_cigar++] = bam1_cigar(b)[k];
- cigar[n_cigar++] = (uint32_t)kl[1]<<4 | BAM_CMATCH; // FIXME: add ref_skip after this line
+ if (bq || zq) {
+ if ((apply_baq && zq) || (!apply_baq && bq)) return -3; // in both cases, do nothing
+ if (bq && apply_baq) { // then convert BQ to ZQ
+ for (i = 0; i < c->l_qseq; ++i)
+ qual[i] = qual[i] + 64 < bq[i]? 0 : qual[i] - ((int)bq[i] - 64);
+ *(bq - 3) = 'Z';
+ } else if (zq && !apply_baq) { // then convert ZQ to BQ
+ for (i = 0; i < c->l_qseq; ++i)
+ qual[i] += (int)zq[i] - 64;
+ *(zq - 3) = 'B';
+ }
+ return 0;
}
- if (q[1] + 1 != c->l_qseq)
- cigar[n_cigar++] = (uint32_t)(c->l_qseq - q[1] - 1)<<4 | BAM_CSOFT_CLIP;
- // copy over the alignment
- if (4 * (n_cigar - (int)c->n_cigar) + b->data_len > b->m_data) { // enlarge b->data
- b->m_data = 4 * (n_cigar - (int)c->n_cigar) + b->data_len;
- kroundup32(b->m_data);
- b->data = realloc(b->data, b->m_data);
+ // find the start and end of the alignment
+ x = c->pos, y = 0, yb = ye = xb = xe = -1;
+ for (k = 0; k < c->n_cigar; ++k) {
+ int op, l;
+ op = cigar[k]&0xf; l = cigar[k]>>4;
+ if (op == BAM_CMATCH) {
+ if (yb < 0) yb = y;
+ if (xb < 0) xb = x;
+ ye = y + l; xe = x + l;
+ x += l; y += l;
+ } else if (op == BAM_CSOFT_CLIP || op == BAM_CINS) y += l;
+ else if (op == BAM_CDEL) x += l;
+ else if (op == BAM_CREF_SKIP) return -1; // do nothing if there is a reference skip
}
- if (n_cigar != (int)c->n_cigar) { // move data
- memmove(b->data + c->l_qname + 4 * n_cigar, bam1_seq(b), b->data_len - c->l_qname - 4 * c->n_cigar);
- b->data_len += 4 * (n_cigar - (int)c->n_cigar);
+ // set bandwidth and the start and the end
+ bw = 7;
+ if (abs((xe - xb) - (ye - yb)) > bw)
+ bw = abs((xe - xb) - (ye - yb)) + 3;
+ conf.bw = bw;
+ xb -= yb + bw/2; if (xb < 0) xb = 0;
+ xe += c->l_qseq - ye + bw/2;
+ if (xe - xb - c->l_qseq > bw)
+ xb += (xe - xb - c->l_qseq - bw) / 2, xe -= (xe - xb - c->l_qseq - bw) / 2;
+ { // glocal
+ uint8_t *s, *r, *q, *seq = bam1_seq(b), *bq;
+ int *state;
+ bq = calloc(c->l_qseq + 1, 1);
+ memcpy(bq, qual, c->l_qseq);
+ s = calloc(c->l_qseq, 1);
+ for (i = 0; i < c->l_qseq; ++i) s[i] = bam_nt16_nt4_table[bam1_seqi(seq, i)];
+ r = calloc(xe - xb, 1);
+ for (i = xb; i < xe; ++i) {
+ if (ref[i] == 0) { xe = i; break; }
+ r[i-xb] = bam_nt16_nt4_table[bam_nt16_table[(int)ref[i]]];
+ }
+ state = calloc(c->l_qseq, sizeof(int));
+ q = calloc(c->l_qseq, 1);
+ kpa_glocal(r, xe-xb, s, c->l_qseq, qual, &conf, state, q);
+ if (!extend_baq) { // in this block, bq[] is capped by base quality qual[]
+ for (k = 0, x = c->pos, y = 0; k < c->n_cigar; ++k) {
+ int op = cigar[k]&0xf, l = cigar[k]>>4;
+ if (op == BAM_CMATCH) {
+ for (i = y; i < y + l; ++i) {
+ if ((state[i]&3) != 0 || state[i]>>2 != x - xb + (i - y)) bq[i] = 0;
+ else bq[i] = bq[i] < q[i]? bq[i] : q[i];
+ }
+ x += l; y += l;
+ } else if (op == BAM_CSOFT_CLIP || op == BAM_CINS) y += l;
+ else if (op == BAM_CDEL) x += l;
+ }
+ for (i = 0; i < c->l_qseq; ++i) bq[i] = qual[i] - bq[i] + 64; // finalize BQ
+ } else { // in this block, bq[] is BAQ that can be larger than qual[] (different from the above!)
+ uint8_t *left, *rght;
+ left = calloc(c->l_qseq, 1); rght = calloc(c->l_qseq, 1);
+ for (k = 0, x = c->pos, y = 0; k < c->n_cigar; ++k) {
+ int op = cigar[k]&0xf, l = cigar[k]>>4;
+ if (op == BAM_CMATCH) {
+ for (i = y; i < y + l; ++i)
+ bq[i] = ((state[i]&3) != 0 || state[i]>>2 != x - xb + (i - y))? 0 : q[i];
+ for (left[y] = bq[y], i = y + 1; i < y + l; ++i)
+ left[i] = bq[i] > left[i-1]? bq[i] : left[i-1];
+ for (rght[y+l-1] = bq[y+l-1], i = y + l - 2; i >= y; --i)
+ rght[i] = bq[i] > rght[i+1]? bq[i] : rght[i+1];
+ for (i = y; i < y + l; ++i)
+ bq[i] = left[i] < rght[i]? left[i] : rght[i];
+ x += l; y += l;
+ } else if (op == BAM_CSOFT_CLIP || op == BAM_CINS) y += l;
+ else if (op == BAM_CDEL) x += l;
+ }
+ for (i = 0; i < c->l_qseq; ++i) bq[i] = 64 + (qual[i] <= bq[i]? 0 : qual[i] - bq[i]); // finalize BQ
+ free(left); free(rght);
+ }
+ if (apply_baq) {
+ for (i = 0; i < c->l_qseq; ++i) qual[i] -= bq[i] - 64; // modify qual
+ bam_aux_append(b, "ZQ", 'Z', c->l_qseq + 1, bq);
+ } else bam_aux_append(b, "BQ", 'Z', c->l_qseq + 1, bq);
+ free(bq); free(s); free(r); free(q); free(state);
}
- memcpy(bam1_cigar(b), cigar, n_cigar * 4);
- c->n_cigar = n_cigar;
- free(cigar);
return 0;
}
+int bam_prob_realn(bam1_t *b, const char *ref)
+{
+ return bam_prob_realn_core(b, ref, 1);
+}
+
int bam_fillmd(int argc, char *argv[])
{
- int c, is_equal = 0, tid = -2, ret, len, is_bam_out, is_sam_in, is_uncompressed, max_nm = 0, is_realn, capQ = 0;
+ int c, is_equal, tid = -2, ret, len, is_bam_out, is_sam_in, is_uncompressed, max_nm, is_realn, capQ, baq_flag;
samfile_t *fp, *fpout = 0;
faidx_t *fai;
char *ref = 0, mode_w[8], mode_r[8];
bam1_t *b;
- is_bam_out = is_sam_in = is_uncompressed = is_realn = 0;
+ is_equal = is_bam_out = is_sam_in = is_uncompressed = is_realn = max_nm = capQ = baq_flag = 0;
mode_w[0] = mode_r[0] = 0;
strcpy(mode_r, "r"); strcpy(mode_w, "w");
- while ((c = getopt(argc, argv, "reubSC:n:")) >= 0) {
+ while ((c = getopt(argc, argv, "EreubSC:n:A")) >= 0) {
switch (c) {
case 'r': is_realn = 1; break;
case 'e': is_equal = 1; break;
case 'S': is_sam_in = 1; break;
case 'n': max_nm = atoi(optarg); break;
case 'C': capQ = atoi(optarg); break;
+ case 'A': baq_flag |= 1; break;
+ case 'E': baq_flag |= 2; break;
default: fprintf(stderr, "[bam_fillmd] unrecognized option '-%c'\n", c); return 1;
}
}
fprintf(stderr, " -u uncompressed BAM output (for piping)\n");
fprintf(stderr, " -b compressed BAM output\n");
fprintf(stderr, " -S the input is SAM with header\n");
- fprintf(stderr, " -r read-independent local realignment\n\n");
+ fprintf(stderr, " -A modify the quality string\n");
+ fprintf(stderr, " -r compute the BQ tag (without -A) or cap baseQ by BAQ (with -A)\n");
+ fprintf(stderr, " -E extended BAQ for better sensitivity but lower specificity\n\n");
return 1;
}
fp = samopen(argv[optind], mode_r, 0);
fprintf(stderr, "[bam_fillmd] fail to find sequence '%s' in the reference.\n",
fp->header->target_name[tid]);
}
- if (is_realn) bam_realn(b, ref);
+ if (is_realn) bam_prob_realn_core(b, ref, baq_flag);
if (capQ > 10) {
int q = bam_cap_mapQ(b, ref, capQ);
if (b->core.qual > q) b->core.qual = q;
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