#include <math.h>
+#include <assert.h>
#include "bam.h"
#include "bam_maqcns.h"
#include "ksort.h"
+#include "errmod.h"
+#include "kaln.h"
KSORT_INIT_GENERIC(uint32_t)
+#define INDEL_WINDOW_SIZE 50
+#define INDEL_EXT_DEP 0.9
+
typedef struct __bmc_aux_t {
int max;
uint32_t *info;
+ uint16_t *info16;
+ errmod_t *em;
} bmc_aux_t;
typedef struct {
float esum[4], fsum[4];
uint32_t c[4];
- uint32_t mapQ_max;
} glf_call_aux_t;
/*
P(<b1,b2>) = \theta \sum_{i=1}^{N-1} 1/i
P(D|<b1,b2>) = \sum_{k=1}^{N-1} p_k 1/2 [(k/N)^n_2(1-k/N)^n_1 + (k/N)^n1(1-k/N)^n_2]
- p_k = i/k / \sum_{i=1}^{N-1} 1/i
+ p_k = 1/k / \sum_{i=1}^{N-1} 1/i
*/
static void cal_het(bam_maqcns_t *aa)
{
int k, n1, n2;
double sum_harmo; // harmonic sum
double poly_rate;
- double p1 = 0.0, p3 = 0.0; // just for testing
free(aa->lhet);
aa->lhet = (double*)calloc(256 * 256, sizeof(double));
for (n1 = 0; n1 < 256; ++n1) {
for (n2 = 0; n2 < 256; ++n2) {
long double sum = 0.0;
- double lC = lgamma(n1+n2+1) - lgamma(n1+1) - lgamma(n2+1); // \binom{n1+n2}{n1}
+ double lC = aa->errmod == BAM_ERRMOD_SOAP? 0 : lgamma(n1+n2+1) - lgamma(n1+1) - lgamma(n2+1);
for (k = 1; k <= aa->n_hap - 1; ++k) {
double pk = 1.0 / k / sum_harmo;
double log1 = log((double)k/aa->n_hap);
sum += pk * 0.5 * (expl(log1*n2) * expl(log2*n1) + expl(log1*n1) * expl(log2*n2));
}
aa->lhet[n1<<8|n2] = lC + logl(sum);
- if (n1 == 17 && n2 == 3) p3 = lC + logl(expl(logl(0.5) * 20));
- if (n1 == 19 && n2 == 1) p1 = lC + logl(expl(logl(0.5) * 20));
}
}
poly_rate = aa->het_rate * sum_harmo;
long double sum_a[257], b[256], q_c[256], tmp[256], fk2[256];
double *lC;
- lC = (double*)calloc(256 * 256, sizeof(double));
+ if (aa->errmod == BAM_ERRMOD_MAQ2) return; // no need to do the following
// aa->lhet will be allocated and initialized
free(aa->fk); free(aa->coef);
+ aa->coef = 0;
aa->fk = (double*)calloc(256, sizeof(double));
- aa->coef = (double*)calloc(256*256*64, sizeof(double));
aa->fk[0] = fk2[0] = 1.0;
for (n = 1; n != 256; ++n) {
aa->fk[n] = pow(aa->theta, n) * (1.0 - aa->eta) + aa->eta;
fk2[n] = aa->fk[n>>1]; // this is an approximation, assuming reads equally likely come from both strands
}
+ if (aa->errmod == BAM_ERRMOD_SOAP) return;
+ aa->coef = (double*)calloc(256*256*64, sizeof(double));
+ lC = (double*)calloc(256 * 256, sizeof(double));
for (n = 1; n != 256; ++n)
for (k = 1; k <= n; ++k)
lC[n<<8|k] = lgamma(n+1) - lgamma(k+1) - lgamma(n-k+1);
bm = (bam_maqcns_t*)calloc(1, sizeof(bam_maqcns_t));
bm->aux = (bmc_aux_t*)calloc(1, sizeof(bmc_aux_t));
bm->het_rate = 0.001;
- bm->theta = 0.85;
+ bm->theta = 0.83f;
bm->n_hap = 2;
bm->eta = 0.03;
+ bm->cap_mapQ = 60;
+ bm->min_baseQ = 13;
return bm;
}
void bam_maqcns_prepare(bam_maqcns_t *bm)
{
+ if (bm->errmod == BAM_ERRMOD_MAQ2) bm->aux->em = errmod_init(1. - bm->theta);
cal_coef(bm); cal_het(bm);
}
void bam_maqcns_destroy(bam_maqcns_t *bm)
{
if (bm == 0) return;
- free(bm->lhet); free(bm->fk); free(bm->coef); free(bm->aux->info);
+ free(bm->lhet); free(bm->fk); free(bm->coef); free(bm->aux->info); free(bm->aux->info16);
+ if (bm->aux->em) errmod_destroy(bm->aux->em);
free(bm->aux); free(bm);
}
glf1_t *bam_maqcns_glfgen(int _n, const bam_pileup1_t *pl, uint8_t ref_base, bam_maqcns_t *bm)
{
- glf_call_aux_t *b;
+ glf_call_aux_t *b = 0;
int i, j, k, w[8], c, n;
glf1_t *g = (glf1_t*)calloc(1, sizeof(glf1_t));
float p[16], min_p = 1e30;
+ uint64_t rms;
g->ref_base = ref_base;
if (_n == 0) return g;
bm->aux->max = _n;
kroundup32(bm->aux->max);
bm->aux->info = (uint32_t*)realloc(bm->aux->info, 4 * bm->aux->max);
+ bm->aux->info16 = (uint16_t*)realloc(bm->aux->info16, 2 * bm->aux->max);
}
- for (i = n = 0; i < _n; ++i) {
+ for (i = n = 0, rms = 0; i < _n; ++i) {
const bam_pileup1_t *p = pl + i;
- uint32_t q, x = 0;
- if (p->is_del || (p->b->core.flag&BAM_FUNMAP)) continue;
+ uint32_t q, x = 0, qq;
+ uint16_t y = 0;
+ if (p->is_del || p->is_refskip || (p->b->core.flag&BAM_FUNMAP)) continue;
q = (uint32_t)bam1_qual(p->b)[p->qpos];
+ if (q < bm->min_baseQ) continue;
x |= (uint32_t)bam1_strand(p->b) << 18 | q << 8 | p->b->core.qual;
+ y |= bam1_strand(p->b)<<4;
if (p->b->core.qual < q) q = p->b->core.qual;
+ c = p->b->core.qual < bm->cap_mapQ? p->b->core.qual : bm->cap_mapQ;
+ rms += c * c;
x |= q << 24;
- q = bam_nt16_nt4_table[bam1_seqi(bam1_seq(p->b), p->qpos)];
- if (!p->is_del && q < 4) x |= 1 << 21 | q << 16;
+ y |= q << 5;
+ qq = bam1_seqi(bam1_seq(p->b), p->qpos);
+ q = bam_nt16_nt4_table[qq? qq : ref_base];
+ if (!p->is_del && !p->is_refskip && q < 4) x |= 1 << 21 | q << 16, y |= q;
+ bm->aux->info16[n] = y;
bm->aux->info[n++] = x;
}
+ rms = (uint8_t)(sqrt((double)rms / n) + .499);
+ if (bm->errmod == BAM_ERRMOD_MAQ2) {
+ errmod_cal(bm->aux->em, n, 4, bm->aux->info16, p);
+ goto goto_glf;
+ }
ks_introsort(uint32_t, n, bm->aux->info);
// generate esum and fsum
b = (glf_call_aux_t*)calloc(1, sizeof(glf_call_aux_t));
for (k = 0; k != 8; ++k) w[k] = 0;
- b->mapQ_max = 0;
for (j = n - 1; j >= 0; --j) { // calculate esum and fsum
uint32_t info = bm->aux->info[j];
if (info>>24 < 4 && (info>>8&0x3f) != 0) info = 4<<24 | (info&0xffffff);
if (w[k] < 0xff) ++w[k];
++b->c[k&3];
}
- if (b->mapQ_max < (info&0x7f)) b->mapQ_max = info&0x7f;
}
// rescale ->c[]
for (j = c = 0; j != 4; ++j) c += b->c[j];
for (j = 0; j != 4; ++j) b->c[j] = (int)(254.0 * b->c[j] / c + 0.5);
for (j = c = 0; j != 4; ++j) c += b->c[j];
}
- // generate likelihood
- for (j = 0; j != 4; ++j) {
- // homozygous
- float tmp1, tmp3;
- int tmp2, bar_e;
- for (k = 0, tmp1 = tmp3 = 0.0, tmp2 = 0; k != 4; ++k) {
- if (j == k) continue;
- tmp1 += b->esum[k]; tmp2 += b->c[k]; tmp3 += b->fsum[k];
- }
- if (tmp2) {
- bar_e = (int)(tmp1 / tmp3 + 0.5);
- if (bar_e < 4) bar_e = 4; // should not happen
- if (bar_e > 63) bar_e = 63;
- p[j<<2|j] = tmp1 + bm->coef[bar_e<<16|c<<8|tmp2];
- } else p[j<<2|j] = 0.0; // all the bases are j
- // heterozygous
- for (k = j + 1; k < 4; ++k) {
- for (i = 0, tmp2 = 0, tmp1 = tmp3 = 0.0; i != 4; ++i) {
- if (i == j || i == k) continue;
- tmp1 += b->esum[i]; tmp2 += b->c[i]; tmp3 += b->fsum[i];
+ if (bm->errmod == BAM_ERRMOD_MAQ) {
+ // generate likelihood
+ for (j = 0; j != 4; ++j) {
+ // homozygous
+ float tmp1, tmp3;
+ int tmp2, bar_e;
+ for (k = 0, tmp1 = tmp3 = 0.0, tmp2 = 0; k != 4; ++k) {
+ if (j == k) continue;
+ tmp1 += b->esum[k]; tmp2 += b->c[k]; tmp3 += b->fsum[k];
}
if (tmp2) {
bar_e = (int)(tmp1 / tmp3 + 0.5);
- if (bar_e < 4) bar_e = 4;
+ if (bar_e < 4) bar_e = 4; // should not happen
if (bar_e > 63) bar_e = 63;
- p[j<<2|k] = p[k<<2|j] = -4.343 * bm->lhet[b->c[j]<<8|b->c[k]] + tmp1 + bm->coef[bar_e<<16|c<<8|tmp2];
- } else p[j<<2|k] = p[k<<2|j] = -4.343 * bm->lhet[b->c[j]<<8|b->c[k]]; // all the bases are either j or k
+ p[j<<2|j] = tmp1 + bm->coef[bar_e<<16|c<<8|tmp2];
+ } else p[j<<2|j] = 0.0; // all the bases are j
+ // heterozygous
+ for (k = j + 1; k < 4; ++k) {
+ for (i = 0, tmp2 = 0, tmp1 = tmp3 = 0.0; i != 4; ++i) {
+ if (i == j || i == k) continue;
+ tmp1 += b->esum[i]; tmp2 += b->c[i]; tmp3 += b->fsum[i];
+ }
+ if (tmp2) {
+ bar_e = (int)(tmp1 / tmp3 + 0.5);
+ if (bar_e < 4) bar_e = 4;
+ if (bar_e > 63) bar_e = 63;
+ p[j<<2|k] = p[k<<2|j] = -4.343 * bm->lhet[b->c[j]<<8|b->c[k]] + tmp1 + bm->coef[bar_e<<16|c<<8|tmp2];
+ } else p[j<<2|k] = p[k<<2|j] = -4.343 * bm->lhet[b->c[j]<<8|b->c[k]]; // all the bases are either j or k
+ }
+ //
+ for (k = 0; k != 4; ++k)
+ if (p[j<<2|k] < 0.0) p[j<<2|k] = 0.0;
+ }
+
+ { // fix p[k<<2|k]
+ float max1, max2, min1, min2;
+ int max_k, min_k;
+ max_k = min_k = -1;
+ max1 = max2 = -1.0; min1 = min2 = 1e30;
+ for (k = 0; k < 4; ++k) {
+ if (b->esum[k] > max1) {
+ max2 = max1; max1 = b->esum[k]; max_k = k;
+ } else if (b->esum[k] > max2) max2 = b->esum[k];
+ }
+ for (k = 0; k < 4; ++k) {
+ if (p[k<<2|k] < min1) {
+ min2 = min1; min1 = p[k<<2|k]; min_k = k;
+ } else if (p[k<<2|k] < min2) min2 = p[k<<2|k];
+ }
+ if (max1 > max2 && (min_k != max_k || min1 + 1.0 > min2))
+ p[max_k<<2|max_k] = min1 > 1.0? min1 - 1.0 : 0.0;
+ }
+ } else if (bm->errmod == BAM_ERRMOD_SOAP) { // apply the SOAP model
+ // generate likelihood
+ for (j = 0; j != 4; ++j) {
+ float tmp;
+ // homozygous
+ for (k = 0, tmp = 0.0; k != 4; ++k)
+ if (j != k) tmp += b->esum[k];
+ p[j<<2|j] = tmp;
+ // heterozygous
+ for (k = j + 1; k < 4; ++k) {
+ for (i = 0, tmp = 0.0; i != 4; ++i)
+ if (i != j && i != k) tmp += b->esum[i];
+ p[j<<2|k] = p[k<<2|j] = -4.343 * bm->lhet[b->c[j]<<8|b->c[k]] + tmp;
+ }
}
- //
- for (k = 0; k != 4; ++k)
- if (p[j<<2|k] < 0.0) p[j<<2|k] = 0.0;
}
+goto_glf:
// convert necessary information to glf1_t
- g->ref_base = ref_base; g->max_mapQ = b->mapQ_max;
+ g->ref_base = ref_base; g->max_mapQ = rms;
g->depth = n > 16777215? 16777215 : n;
for (j = 0; j != 4; ++j)
for (k = j; k < 4; ++k)
uint32_t glf2cns(const glf1_t *g, int q_r)
{
- int i, j, k, tmp[16], min = 10000, min2 = 10000, min3 = 10000, min_g = -1, min_g2 = -1;
+ int i, j, k, p[10], ref4;
uint32_t x = 0;
+ ref4 = bam_nt16_nt4_table[g->ref_base];
for (i = k = 0; i < 4; ++i)
for (j = i; j < 4; ++j) {
- tmp[j<<2|i] = -1;
- tmp[i<<2|j] = g->lk[k++] + (i == j? 0 : q_r);
+ int prior = (i == ref4 && j == ref4? 0 : i == ref4 || j == ref4? q_r : q_r + 3);
+ p[k] = (g->lk[k] + prior)<<4 | i<<2 | j;
+ ++k;
}
- for (i = 0; i < 16; ++i) {
- if (tmp[i] < 0) continue;
- if (tmp[i] < min) {
- min3 = min2; min2 = min; min = tmp[i]; min_g2 = min_g; min_g = i;
- } else if (tmp[i] < min2) {
- min3 = min2; min2 = tmp[i]; min_g2 = i;
- } else if (tmp[i] < min3) min3 = tmp[i];
- }
- x = min_g >= 0? (1U<<(min_g>>2&3) | 1U<<(min_g&3)) << 28 : 0xf << 28;
- x |= min_g2 >= 0? (1U<<(min_g2>>2&3) | 1U<<(min_g2&3)) << 24 : 0xf << 24;
- x |= (uint32_t)g->max_mapQ << 16;
- x |= min2 < 10000? (min2 - min < 256? min2 - min : 255) << 8 : 0xff << 8;
- x |= min2 < 10000 && min3 < 10000? (min3 - min2 < 256? min3 - min2 : 255) : 0xff;
+ for (i = 1; i < 10; ++i) // insertion sort
+ for (j = i; j > 0 && p[j] < p[j-1]; --j)
+ k = p[j], p[j] = p[j-1], p[j-1] = k;
+ x = (1u<<(p[0]&3) | 1u<<(p[0]>>2&3)) << 28; // the best genotype
+ x |= (uint32_t)g->max_mapQ << 16; // rms mapQ
+ x |= ((p[1]>>4) - (p[0]>>4) < 256? (p[1]>>4) - (p[0]>>4) : 255) << 8; // consensus Q
+ for (k = 0; k < 10; ++k)
+ if ((p[k]&0xf) == (ref4<<2|ref4)) break;
+ if (k == 10) k = 9;
+ x |= (p[k]>>4) - (p[0]>>4) < 256? (p[k]>>4) - (p[0]>>4) : 255; // snp Q
return x;
}
uint32_t x;
if (n) {
g = bam_maqcns_glfgen(n, pl, 0xf, bm);
- x = glf2cns(g, (int)(bm->q_r + 0.5));
+ x = g->depth == 0? (0xfU<<28 | 0xfU<<24) : glf2cns(g, (int)(bm->q_r + 0.5));
free(g);
} else x = 0xfU<<28 | 0xfU<<24;
return x;
bam_maqindel_opt_t *bam_maqindel_opt_init()
{
bam_maqindel_opt_t *mi = (bam_maqindel_opt_t*)calloc(1, sizeof(bam_maqindel_opt_t));
+ mi->q_indel = 40;
+ mi->r_indel = 0.00015;
+ mi->r_snp = 0.001;
+ //
mi->mm_penalty = 3;
mi->indel_err = 4;
mi->ambi_thres = 10;
void bam_maqindel_ret_destroy(bam_maqindel_ret_t *mir)
{
if (mir == 0) return;
- free(mir->s1); free(mir->s2); free(mir);
+ free(mir->s[0]); free(mir->s[1]); free(mir);
+}
+
+int bam_tpos2qpos(const bam1_core_t *c, const uint32_t *cigar, int32_t tpos, int is_left, int32_t *_tpos)
+{
+ int k, x = c->pos, y = 0, last_y = 0;
+ *_tpos = c->pos;
+ for (k = 0; k < c->n_cigar; ++k) {
+ int op = cigar[k] & BAM_CIGAR_MASK;
+ int l = cigar[k] >> BAM_CIGAR_SHIFT;
+ if (op == BAM_CMATCH) {
+ if (c->pos > tpos) return y;
+ if (x + l > tpos) {
+ *_tpos = tpos;
+ return y + (tpos - x);
+ }
+ x += l; y += l;
+ last_y = y;
+ } else if (op == BAM_CINS || op == BAM_CSOFT_CLIP) y += l;
+ else if (op == BAM_CDEL || op == BAM_CREF_SKIP) {
+ if (x + l > tpos) {
+ *_tpos = is_left? x : x + l;
+ return y;
+ }
+ x += l;
+ }
+ }
+ *_tpos = x;
+ return last_y;
}
#define MINUS_CONST 0x10000000
-bam_maqindel_ret_t *bam_maqindel(int n, int pos, const bam_maqindel_opt_t *mi, const bam_pileup1_t *pl, const char *ref)
+bam_maqindel_ret_t *bam_maqindel(int n, int pos, const bam_maqindel_opt_t *mi, const bam_pileup1_t *pl, const char *ref,
+ int _n_types, int *_types)
{
- int i, j, n_types, *types, left, right;
+ int i, j, n_types, *types, left, right, max_rd_len = 0;
bam_maqindel_ret_t *ret = 0;
- for (i = 0; i < n; ++i) {
- const bam_pileup1_t *p = pl + i;
- if (!(p->b->core.flag&BAM_FUNMAP) && p->indel != 0) break;
+ // if there is no proposed indel, check if there is an indel from the alignment
+ if (_n_types == 0) {
+ for (i = 0; i < n; ++i) {
+ const bam_pileup1_t *p = pl + i;
+ if (!(p->b->core.flag&BAM_FUNMAP) && p->indel != 0) break;
+ }
+ if (i == n) return 0; // no indel
}
- if (i == n) return 0; // no indel
{ // calculate how many types of indels are available (set n_types and types)
int m;
uint32_t *aux;
- aux = (uint32_t*)calloc(n+1, 4);
+ aux = (uint32_t*)calloc(n + _n_types + 1, 4);
m = 0;
aux[m++] = MINUS_CONST; // zero indel is always a type
for (i = 0; i < n; ++i) {
const bam_pileup1_t *p = pl + i;
if (!(p->b->core.flag&BAM_FUNMAP) && p->indel != 0)
aux[m++] = MINUS_CONST + p->indel;
+ j = bam_cigar2qlen(&p->b->core, bam1_cigar(p->b));
+ if (j > max_rd_len) max_rd_len = j;
}
+ if (_n_types) // then also add this to aux[]
+ for (i = 0; i < _n_types; ++i)
+ if (_types[i]) aux[m++] = MINUS_CONST + _types[i];
ks_introsort(uint32_t, m, aux);
- n_types = 1;
- for (i = 1; i < m; ++i)
+ // squeeze out identical types
+ for (i = 1, n_types = 1; i < m; ++i)
if (aux[i] != aux[i-1]) ++n_types;
types = (int*)calloc(n_types, sizeof(int));
j = 0;
free(aux);
}
{ // calculate left and right boundary
- bam_segreg_t seg;
- left = 0x7fffffff; right = 0;
- for (i = 0; i < n; ++i) {
- const bam_pileup1_t *p = pl + i;
- if (!(p->b->core.flag&BAM_FUNMAP)) {
- bam_segreg(pos, &p->b->core, bam1_cigar(p->b), &seg);
- if (seg.tbeg < left) left = seg.tbeg;
- if (seg.tend > right) right = seg.tend;
- }
- }
+ left = pos > INDEL_WINDOW_SIZE? pos - INDEL_WINDOW_SIZE : 0;
+ right = pos + INDEL_WINDOW_SIZE;
+ if (types[0] < 0) right -= types[0];
+ // in case the alignments stand out the reference
+ for (i = pos; i < right; ++i)
+ if (ref[i] == 0) break;
+ right = i;
}
{ // the core part
- char *ref2, *inscns = 0;
- int k, l, *score, max_ins = types[n_types-1];
- ref2 = (char*)calloc(right - left + types[n_types-1] + 2, 1);
+ char *ref2, *rs, *inscns = 0;
+ int qr_snp, k, l, *score, *pscore, max_ins = types[n_types-1];
+ qr_snp = (int)(-4.343 * log(mi->r_snp) + .499);
if (max_ins > 0) { // get the consensus of inserted sequences
int *inscns_aux = (int*)calloc(4 * n_types * max_ins, sizeof(int));
// count occurrences
}
}
}
- // construct the consensus
+ // construct the consensus of inserted sequence
inscns = (char*)calloc(n_types * max_ins, sizeof(char));
for (i = 0; i < n_types; ++i) {
for (j = 0; j < types[i]; ++j) {
free(inscns_aux);
}
// calculate score
+ ref2 = (char*)calloc(right - left + types[n_types-1] + 2, 1);
+ rs = (char*)calloc(right - left + max_rd_len + types[n_types-1] + 2, 1);
score = (int*)calloc(n_types * n, sizeof(int));
+ pscore = (int*)calloc(n_types * n, sizeof(int));
for (i = 0; i < n_types; ++i) {
+ ka_param_t ap = ka_param_blast;
+ ap.band_width = 2 * types[n_types - 1] + 2;
+ ap.gap_end_ext = 0;
// write ref2
for (k = 0, j = left; j <= pos; ++j)
- ref2[k++] = bam_nt16_table[(int)ref[j]];
+ ref2[k++] = bam_nt16_nt4_table[bam_nt16_table[(int)ref[j]]];
if (types[i] <= 0) j += -types[i];
else for (l = 0; l < types[i]; ++l)
- ref2[k++] = inscns[i*max_ins + l];
+ ref2[k++] = bam_nt16_nt4_table[(int)inscns[i*max_ins + l]];
+ if (types[0] < 0) { // mask deleted sequences
+ int jj, tmp = types[i] >= 0? -types[0] : -types[0] + types[i];
+ for (jj = 0; jj < tmp && j < right && ref[j]; ++jj, ++j)
+ ref2[k++] = 4;
+ }
for (; j < right && ref[j]; ++j)
- ref2[k++] = bam_nt16_table[(int)ref[j]];
+ ref2[k++] = bam_nt16_nt4_table[bam_nt16_table[(int)ref[j]]];
+ if (j < right) right = j;
// calculate score for each read
for (j = 0; j < n; ++j) {
const bam_pileup1_t *p = pl + j;
- uint32_t *cigar;
- bam1_core_t *c = &p->b->core;
- int s;
- bam_segreg_t seg;
- if (c->flag&BAM_FUNMAP) continue;
- cigar = bam1_cigar(p->b);
- bam_segreg(pos, c, cigar, &seg);
- for (s = 0, l = seg.qbeg; c->pos + l < right && l < seg.qend; ++l) {
- int cq = bam1_seqi(bam1_seq(p->b), l), ct;
- ct = c->pos + l >= left? ref2[c->pos + l - left] : 15; // "<" should not happen if there is no bug
- if (cq < 15 && ct < 15)
- s += cq == ct? 1 : -mi->mm_penalty;
- }
- score[i*n + j] = s;
- if (types[i] != 0) { // then try the other way to calculate the score
- for (s = 0, l = seg.qbeg; c->pos + l + types[i] < right && l < seg.qend; ++l) {
- int cq = bam1_seqi(bam1_seq(p->b), l), ct;
- ct = c->pos + l + types[i] >= left? ref2[c->pos + l + types[i] - left] : 15;
- if (cq < 15 && ct < 15)
- s += cq == ct? 1 : -mi->mm_penalty;
+ int qbeg, qend, tbeg, tend;
+ if (p->b->core.flag & BAM_FUNMAP) continue;
+ qbeg = bam_tpos2qpos(&p->b->core, bam1_cigar(p->b), left, 0, &tbeg);
+ qend = bam_tpos2qpos(&p->b->core, bam1_cigar(p->b), right, 1, &tend);
+ assert(tbeg >= left);
+ for (l = qbeg; l < qend; ++l)
+ rs[l - qbeg] = bam_nt16_nt4_table[bam1_seqi(bam1_seq(p->b), l)];
+ {
+ int x, y, n_acigar, ps;
+ uint32_t *acigar;
+ ps = 0;
+ if (tend - tbeg + types[i] <= 0) {
+ score[i*n+j] = -(1<<20);
+ pscore[i*n+j] = 1<<20;
+ continue;
+ }
+ acigar = ka_global_core((uint8_t*)ref2 + tbeg - left, tend - tbeg + types[i], (uint8_t*)rs, qend - qbeg, &ap, &score[i*n+j], &n_acigar);
+ x = tbeg - left; y = 0;
+ for (l = 0; l < n_acigar; ++l) {
+ int op = acigar[l]&0xf;
+ int len = acigar[l]>>4;
+ if (op == BAM_CMATCH) {
+ int k;
+ for (k = 0; k < len; ++k)
+ if (ref2[x+k] != rs[y+k] && ref2[x+k] < 4)
+ ps += bam1_qual(p->b)[y+k] < qr_snp? bam1_qual(p->b)[y+k] : qr_snp;
+ x += len; y += len;
+ } else if (op == BAM_CINS || op == BAM_CSOFT_CLIP) {
+ if (op == BAM_CINS && l > 0 && l < n_acigar - 1) ps += mi->q_indel * len;
+ y += len;
+ } else if (op == BAM_CDEL) {
+ if (l > 0 && l < n_acigar - 1) ps += mi->q_indel * len;
+ x += len;
+ }
}
+ pscore[i*n+j] = ps;
+ /*if (1) { // for debugging only
+ fprintf(stderr, "id=%d, pos=%d, type=%d, j=%d, score=%d, psore=%d, %d, %d, %d, %d, %d, ",
+ j, pos+1, types[i], j, score[i*n+j], pscore[i*n+j], tbeg, tend, qbeg, qend, mi->q_indel);
+ for (l = 0; l < n_acigar; ++l) fprintf(stderr, "%d%c", acigar[l]>>4, "MIDS"[acigar[l]&0xf]);
+ fprintf(stderr, "\n");
+ for (l = 0; l < tend - tbeg + types[i]; ++l) fputc("ACGTN"[ref2[l+tbeg-left]], stderr);
+ fputc('\n', stderr);
+ for (l = 0; l < qend - qbeg; ++l) fputc("ACGTN"[rs[l]], stderr);
+ fputc('\n', stderr);
+ }*/
+ free(acigar);
}
- if (score[i*n+j] < s) score[i*n+j] = s; // choose the higher of the two scores
- if (types[i] != 0) score[i*n+j] -= mi->indel_err;
- //printf("%d, %d, %d, %d\n", i, types[i], j, score[i*n+j]);
}
}
{ // get final result
sum = (int*)calloc(n_types, sizeof(int));
for (i = 0; i < n_types; ++i)
for (j = 0; j < n; ++j)
- sum[i] += score[i*n+j];
+ sum[i] += -pscore[i*n+j];
max1 = max2 = -0x7fffffff; max1_i = max2_i = -1;
for (i = 0; i < n_types; ++i) {
if (sum[i] > max1) {
// write ret
ret = (bam_maqindel_ret_t*)calloc(1, sizeof(bam_maqindel_ret_t));
ret->indel1 = types[max1_i]; ret->indel2 = types[max2_i];
- ret->s1 = (char*)calloc(abs(ret->indel1) + 2, 1);
- ret->s2 = (char*)calloc(abs(ret->indel2) + 2, 1);
+ ret->s[0] = (char*)calloc(abs(ret->indel1) + 2, 1);
+ ret->s[1] = (char*)calloc(abs(ret->indel2) + 2, 1);
+ // write indel sequence
if (ret->indel1 > 0) {
- ret->s1[0] = '+';
+ ret->s[0][0] = '+';
for (k = 0; k < ret->indel1; ++k)
- ret->s1[k+1] = bam_nt16_rev_table[(int)inscns[max1_i*max_ins + k]];
+ ret->s[0][k+1] = bam_nt16_rev_table[(int)inscns[max1_i*max_ins + k]];
} else if (ret->indel1 < 0) {
- ret->s1[0] = '-';
+ ret->s[0][0] = '-';
for (k = 0; k < -ret->indel1 && ref[pos + k + 1]; ++k)
- ret->s1[k+1] = ref[pos + k + 1];
- } else ret->s1[0] = '*';
+ ret->s[0][k+1] = ref[pos + k + 1];
+ } else ret->s[0][0] = '*';
if (ret->indel2 > 0) {
- ret->s2[0] = '+';
+ ret->s[1][0] = '+';
for (k = 0; k < ret->indel2; ++k)
- ret->s2[k+1] = bam_nt16_rev_table[(int)inscns[max2_i*max_ins + k]];
+ ret->s[1][k+1] = bam_nt16_rev_table[(int)inscns[max2_i*max_ins + k]];
} else if (ret->indel2 < 0) {
- ret->s2[0] = '-';
+ ret->s[1][0] = '-';
for (k = 0; k < -ret->indel2 && ref[pos + k + 1]; ++k)
- ret->s2[k+1] = ref[pos + k + 1];
- } else ret->s2[0] = '*';
- for (j = 0; j < n; ++j) {
- if (score[max1_i*n+j] < 0 && score[max2_i*n+j] < 0) ++ret->cnt_anti;
- else {
- int diff = score[max1_i*n+j] - score[max2_i*n+j];
- if (diff > mi->ambi_thres) ++ret->cnt1;
- else if (diff < -mi->ambi_thres) ++ret->cnt2;
- else ++ret->cnt_ambi;
+ ret->s[1][k+1] = ref[pos + k + 1];
+ } else ret->s[1][0] = '*';
+ // write count
+ for (i = 0; i < n; ++i) {
+ const bam_pileup1_t *p = pl + i;
+ if (p->indel == ret->indel1) ++ret->cnt1;
+ else if (p->indel == ret->indel2) ++ret->cnt2;
+ else ++ret->cnt_anti;
+ }
+ { // write gl[]
+ int tmp, seq_err = 0;
+ double x = 1.0;
+ tmp = max1_i - max2_i;
+ if (tmp < 0) tmp = -tmp;
+ for (j = 0; j < tmp + 1; ++j) x *= INDEL_EXT_DEP;
+ seq_err = mi->q_indel * (1.0 - x) / (1.0 - INDEL_EXT_DEP);
+ ret->gl[0] = ret->gl[1] = 0;
+ for (j = 0; j < n; ++j) {
+ int s1 = pscore[max1_i*n + j], s2 = pscore[max2_i*n + j];
+ //fprintf(stderr, "id=%d, %d, %d, %d, %d, %d\n", j, pl[j].b->core.pos+1, types[max1_i], types[max2_i], s1, s2);
+ if (s1 > s2) ret->gl[0] += s1 - s2 < seq_err? s1 - s2 : seq_err;
+ else ret->gl[1] += s2 - s1 < seq_err? s2 - s1 : seq_err;
}
}
+ // write cnt_ref and cnt_ambi
+ if (max1_i != 0 && max2_i != 0) {
+ for (j = 0; j < n; ++j) {
+ int diff1 = score[j] - score[max1_i * n + j];
+ int diff2 = score[j] - score[max2_i * n + j];
+ if (diff1 > 0 && diff2 > 0) ++ret->cnt_ref;
+ else if (diff1 == 0 || diff2 == 0) ++ret->cnt_ambi;
+ }
+ }
+ }
+ free(score); free(pscore); free(ref2); free(rs); free(inscns);
+ }
+ { // call genotype
+ int q[3], qr_indel = (int)(-4.343 * log(mi->r_indel) + 0.5);
+ int min1, min2, min1_i;
+ q[0] = ret->gl[0] + (ret->s[0][0] != '*'? 0 : 0) * qr_indel;
+ q[1] = ret->gl[1] + (ret->s[1][0] != '*'? 0 : 0) * qr_indel;
+ q[2] = n * 3 + (ret->s[0][0] == '*' || ret->s[1][0] == '*'? 1 : 1) * qr_indel;
+ min1 = min2 = 0x7fffffff; min1_i = -1;
+ for (i = 0; i < 3; ++i) {
+ if (q[i] < min1) {
+ min2 = min1; min1 = q[i]; min1_i = i;
+ } else if (q[i] < min2) min2 = q[i];
}
- free(score); free(ref2); free(inscns);
+ ret->gt = min1_i;
+ ret->q_cns = min2 - min1;
+ // set q_ref
+ if (ret->gt < 2) ret->q_ref = (ret->s[ret->gt][0] == '*')? 0 : q[1-ret->gt] - q[ret->gt] - qr_indel - 3;
+ else ret->q_ref = (ret->s[0][0] == '*')? q[0] - q[2] : q[1] - q[2];
+ if (ret->q_ref < 0) ret->q_ref = 0;
}
free(types);
return ret;