int main_vcfview(int argc, char *argv[])
{
int c, clevel = -1, in_type = FT_BCF, out_type = FT_VCF;
char *fname_out = NULL, moder[8], modew[8];
while ((c = getopt(argc, argv, "l:bvo:n:z?hu")) >= 0) {
switch (c) {
case 'o':
switch (optarg[0]) {
case 'b': out_type = FT_BCF_GZ; break;
case 'u': out_type = FT_BCF; break;
case 'z': out_type = FT_VCF_GZ; break;
case 'v': out_type = FT_VCF; break;
default: error("The output type \"%s\" not recognised\n", optarg);
}
break;
case 'l': clevel = atoi(optarg); out_type |= FT_GZ; break;
case 'v': in_type = FT_VCF; break;
case 'b': out_type = FT_BCF_GZ; break;
case 'u': out_type = FT_BCF; break;
case 'z': out_type = FT_VCF_GZ; break;
case 'n': fname_out = optarg; break;
case '?':
case 'h': usage(); return 1; break;
}
}
if (argc!=optind+1) { usage(); return 1; }
// Init reader
strcpy(moder, "r");
if ( (!strcmp("-",argv[optind]) && (in_type & FT_BCF)) || (hts_file_type(argv[optind]) & FT_BCF)) strcat(moder, "b");
htsFile *fp_in = hts_open(argv[optind], moder, NULL);
if ( !fp_in ) error("Fail to open: %s\n", argv[optind]);
bcf_hdr_t *hdr = vcf_hdr_read(fp_in);
if ( !hdr ) error("Fail to read VCF/BCF header: %s\n", argv[optind]);
bcf1_t *rec = bcf_init1();
// Init writer
strcpy(modew, "w");
if (clevel >= 0 && clevel <= 9) sprintf(modew + 1, "%d", clevel);
if (out_type & FT_GZ) strcat(modew,"z");
if (out_type & FT_BCF) strcat(modew, "b");
if (out_type == FT_BCF) strcat(modew, "u"); // uncompressed BCF output
htsFile *fp_out = hts_open(fname_out ? fname_out : "-", modew, NULL);
vcf_hdr_write(fp_out, hdr);
while ( vcf_read1(fp_in, hdr, rec) >= 0) vcf_write1(fp_out, hdr, rec);
bcf_destroy1(rec);
bcf_hdr_destroy(hdr);
hts_close(fp_in);
hts_close(fp_out);
return 0;
}
static void init_missed_line(args_t *args)
{
int i;
for (i=0; i<bcf_hdr_nsamples(args->aux.hdr); i++)
{
args->aux.gts[i*2] = bcf_gt_missing;
args->aux.gts[i*2+1] = bcf_int32_vector_end;
}
args->missed_line = bcf_init1();
bcf_update_genotypes(args->aux.hdr, args->missed_line, args->aux.gts, 2*bcf_hdr_nsamples(args->aux.hdr));
bcf_float_set_missing(args->missed_line->qual);
}
Variant VariantBuilder::build() const {
const auto new_variant_body = utils::make_shared_variant(bcf_init1());
// Always build from scratch for now, until support for building from a starting variant is added
build_from_scratch(new_variant_body);
if ( m_enable_validation ) {
post_build_validation(new_variant_body);
}
return Variant{m_header.m_header, new_variant_body};
}
/**
* Gets record from pool, creates a new record if necessary
*/
bcf1_t* BCFOrderedReader::get_bcf1_from_pool()
{
if(!pool.empty())
{
bcf1_t* v = pool.front();
pool.pop_front();
return v;
}
else
{
return bcf_init1();
}
};
static void unread_vcf_line(args_t *args, bcf1_t **rec_ptr)
{
bcf1_t *rec = *rec_ptr;
if ( args->vcf_rbuf.n >= args->vcf_rbuf.m )
error("FIXME: too many overlapping records near %s:%d\n", bcf_seqname(args->hdr,rec),rec->pos+1);
// Insert the new record in the buffer. The line would be overwritten in
// the next bcf_sr_next_line call, therefore we need to swap it with an
// unused one
int i = rbuf_append(&args->vcf_rbuf);
if ( !args->vcf_buf[i] ) args->vcf_buf[i] = bcf_init1();
bcf1_t *tmp = rec; *rec_ptr = args->vcf_buf[i]; args->vcf_buf[i] = tmp;
}
static void phased_push(args_t *args, bcf1_t *arec, bcf1_t *brec)
{
if ( arec && arec->errcode )
error("Parse error at %s:%d, cannot proceed: %s\n", bcf_seqname(args->files->readers[0].header,arec),arec->pos+1, args->files->readers[0].fname);
if ( brec && brec->errcode )
error("Parse error at %s:%d, cannot proceed: %s\n", bcf_seqname(args->files->readers[1].header,brec),brec->pos+1, args->files->readers[1].fname);
int i, nsmpl = bcf_hdr_nsamples(args->out_hdr);
int chr_id = bcf_hdr_name2id(args->out_hdr, bcf_seqname(args->files->readers[0].header,arec));
if ( args->prev_chr<0 || args->prev_chr!=chr_id )
{
if ( args->prev_chr>=0 ) phased_flush(args);
for (i=0; i<nsmpl; i++)
args->phase_set[i] = arec->pos+1;
args->phase_set_changed = 1;
if ( args->seen_seq[chr_id] ) error("The chromosome block %s is not contiguous\n", bcf_seqname(args->files->readers[0].header,arec));
args->seen_seq[chr_id] = 1;
args->prev_chr = chr_id;
args->prev_pos_check = -1;
}
if ( !brec )
{
bcf_translate(args->out_hdr, args->files->readers[0].header, arec);
if ( args->nswap )
phase_update(args, args->out_hdr, arec);
if ( !args->compact_PS || args->phase_set_changed )
{
bcf_update_format_int32(args->out_hdr,arec,"PS",args->phase_set,nsmpl);
args->phase_set_changed = 0;
}
bcf_write(args->out_fh, args->out_hdr, arec);
if ( arec->pos < args->prev_pos_check )
error("FIXME, disorder: %s:%d in %s vs %d written [3]\n", bcf_seqname(args->files->readers[0].header,arec), arec->pos+1,args->files->readers[0].fname, args->prev_pos_check+1);
args->prev_pos_check = arec->pos;
return;
}
int m = args->mbuf;
args->nbuf += 2;
hts_expand(bcf1_t*,args->nbuf,args->mbuf,args->buf);
for (i=m; i<args->mbuf; i++)
args->buf[i] = bcf_init1();
SWAP(bcf1_t*, args->files->readers[0].buffer[0], args->buf[args->nbuf-2]);
SWAP(bcf1_t*, args->files->readers[1].buffer[0], args->buf[args->nbuf-1]);
}
bcf1_t *vcfbuf_push(vcfbuf_t *buf, bcf1_t *rec, int swap)
{
if ( !swap ) error("todo: swap=%d\n", swap);
rbuf_expand0(&buf->rbuf, vcfrec_t, buf->rbuf.n+1, buf->vcf);
int i = rbuf_append(&buf->rbuf);
if ( !buf->vcf[i].rec ) buf->vcf[i].rec = bcf_init1();
bcf1_t *ret = buf->vcf[i].rec;
buf->vcf[i].rec = rec;
buf->vcf[i].af_set = 0;
return ret;
}
/**
* Gets record from pool, creates a new record if necessary
*/
bcf1_t* BCFSyncedStreamReader::get_bcf1_from_pool()
{
if(!pool.empty())
{
bcf1_t* v = pool.front();
pool.pop_front();
bcf_clear(v);
return v;
}
else
{
bcf1_t* v = bcf_init1();
bcf_clear(v);
return v;
}
}
int main_getalt(int argc, char *argv[])
{
int c;
char *fn;
BGZF *fp;
bcf1_t *b;
bcf_hdr_t *h;
kstring_t s = {0,0,0};
while ((c = getopt(argc, argv, "")) >= 0) {
}
if (argc - optind == 0) {
fprintf(stderr, "Usage: bgt getalt <bgt-base>\n");
return 1;
}
fn = (char*)calloc(strlen(argv[optind]) + 5, 1);
sprintf(fn, "%s.bcf", argv[optind]);
fp = bgzf_open(fn, "r");
free(fn);
assert(fp);
h = bcf_hdr_read(fp);
b = bcf_init1();
while (bcf_read1(fp, b) >= 0) {
char *ref, *alt;
int l_ref, l_alt, i, min_l;
bcf_get_ref_alt1(b, &l_ref, &ref, &l_alt, &alt);
min_l = l_ref < l_alt? l_ref : l_alt;
for (i = 0; i < min_l && ref[i] == alt[i]; ++i);
s.l = 0;
kputs(h->id[BCF_DT_CTG][b->rid].key, &s);
kputc(':', &s); kputw(b->pos + 1 + i, &s);
kputc(':', &s); kputw(b->rlen - i, &s);
kputc(':', &s); kputsn(alt + i, l_alt - i, &s);
puts(s.s);
}
bcf_destroy1(b);
bcf_hdr_destroy(h);
bgzf_close(fp);
free(s.s);
return 0;
}
static bcf1_t *rec_set_info(args_t *args, bcf1_t *rec)
{
bcf1_t *out = bcf_init1();
out->rid = rec->rid;
out->pos = rec->pos;
out->rlen = rec->rlen;
out->qual = rec->qual;
out->n_allele = rec->n_allele;
out->n_sample = 1;
if ( args->keep_info )
{
out->n_info = rec->n_info;
out->shared.m = out->shared.l = rec->shared.l;
out->shared.s = (char*) malloc(out->shared.l);
memcpy(out->shared.s,rec->shared.s,out->shared.l);
return out;
}
// build the BCF record
kstring_t tmp = {0,0,0};
char *ptr = rec->shared.s;
kputsn_(ptr, rec->unpack_size[0], &tmp); ptr += rec->unpack_size[0]; // ID
kputsn_(ptr, rec->unpack_size[1], &tmp); ptr += rec->unpack_size[1]; // REF+ALT
kputsn_(ptr, rec->unpack_size[2], &tmp); // FILTER
if ( args->ninfo_tags )
{
int i;
for (i=0; i<rec->n_info; i++)
{
bcf_info_t *info = &rec->d.info[i];
int id = info->key;
if ( !args->info_tags[id] ) continue;
kputsn_(info->vptr - info->vptr_off, info->vptr_len + info->vptr_off, &tmp);
out->n_info++;
}
}
out->shared.m = tmp.m;
out->shared.s = tmp.s;
out->shared.l = tmp.l;
out->unpacked = 0;
return out;
}
int main(int argc, char **argv)
{
if (argc < 3) {
fprintf(stderr,"%s <bcf file> <num vars>\n", argv[0]);
return 1;
}
char *fname = argv[1];
uint32_t num_vars = atoi(argv[2]);
htsFile *fp = hts_open(fname,"rb");
bcf_hdr_t *hdr = bcf_hdr_read(fp);
bcf1_t *line = bcf_init1();
int32_t *gt_p = NULL;
uint32_t num_inds = bcf_hdr_nsamples(hdr);
int32_t i, j, k, ntmp = 0, int_i = 0, two_bit_i = 0, sum, t_sum = 0;
uint32_t num_ind_ints = 1 + ((num_inds - 1) / 16);
pri_queue q = priq_new(0);
priority p;
uint32_t *packed_ints = (uint32_t *) calloc(num_ind_ints,
sizeof(uint32_t));
FILE *gt_of = fopen("gt.tmp.packed","wb");
FILE *md_of = fopen("md.tmp.packed","w");
uint32_t *md_index = (uint32_t *) malloc(num_vars * sizeof(uint32_t));
uint32_t md_i = 0;
unsigned long t_bcf_read = 0,
t_bcf_dup = 0,
t_bcf_unpack = 0,
t_bcf_get_genotypes = 0,
t_bcf_hdr_nsamples = 0,
t_q = 0,
t_write = 0,
t_get_md = 0,
t_md_write = 0,
t_pack = 0;
for (i = 0; i < num_vars; ++i) {
sum = 0;
int_i = 0;
two_bit_i = 0;
int r = bcf_read(fp, hdr, line);
// Copy
bcf1_t *t_line = bcf_dup(line);
// Unpack
bcf_unpack(t_line, BCF_UN_ALL);
// Get metadata
size_t len = strlen(bcf_hdr_id2name(hdr, t_line->rid)) +
10 + // max length of pos
strlen(t_line->d.id) +
strlen(t_line->d.allele[0]) +
strlen(t_line->d.allele[1]) +
4; //tabs
char *md = (char *) malloc(len * sizeof(char));
sprintf(md, "%s\t%d\t%s\t%s\t%s",
bcf_hdr_id2name(hdr, t_line->rid),
t_line->pos + 1,
t_line->d.id,
t_line->d.allele[0],
t_line->d.allele[1]);
// Write metadata
md_i += strlen(md);
md_index[i] = md_i;
fprintf(md_of, "%s", md);
// Get gentotypes
uint32_t num_gts_per_sample = bcf_get_genotypes(hdr,
t_line,
>_p,
&ntmp);
num_gts_per_sample /= num_inds;
int32_t *gt_i = gt_p;
// Pack genotypes
for (j = 0; j < num_inds; ++j) {
uint32_t gt = 0;
for (k = 0; k < num_gts_per_sample; ++k) {
gt += bcf_gt_allele(gt_i[k]);
}
packed_ints[int_i] += gt << (30 - 2*two_bit_i);
two_bit_i += 1;
if (two_bit_i == 16) {
two_bit_i = 0;
int_i += 1;
}
sum += gt;
gt_i += num_gts_per_sample;
}
// Get a priority for the variant based on the sum and number of
// leading zeros
p.sum = sum;
uint32_t prefix_len = 0;
j = 0;
while ((j < num_ind_ints) && (packed_ints[j] == 0)){
prefix_len += 32;
j += 1;
}
if (j < num_ind_ints)
prefix_len += nlz1(packed_ints[j]);
// Push it into the q
p.len = prefix_len;
int *j = (int *) malloc (sizeof(int));
j[0] = i;
priq_push(q, j, p);
// Write to file
fwrite(packed_ints, sizeof(uint32_t), num_ind_ints,gt_of);
memset(packed_ints, 0, num_ind_ints*sizeof(uint32_t));
t_sum += sum;
bcf_destroy(t_line);
free(md);
}
fclose(gt_of);
fclose(md_of);
md_of = fopen("md.tmp.packed","r");
FILE *md_out = fopen("md.bim","w");
gt_of = fopen("gt.tmp.packed","rb");
FILE *s_gt_of = fopen("s.gt.tmp.packed","wb");
// Get variants in order and rewrite a variant-major sorted matrix
while ( priq_top(q, &p) != NULL ) {
int *d = priq_pop(q, &p);
uint32_t start = 0;
if (*d != 0)
start = md_index[*d - 1];
uint32_t len = md_index[*d] - start;
fseek(md_of, start*sizeof(char), SEEK_SET);
char buf[len+1];
fread(buf, sizeof(char), len, md_of);
buf[len] = '\0';
fseek(gt_of, (*d)*num_ind_ints*sizeof(uint32_t), SEEK_SET);
fread(packed_ints, sizeof(uint32_t), num_ind_ints, gt_of);
fwrite(packed_ints, sizeof(uint32_t), num_ind_ints,s_gt_of);
fprintf(md_out, "%s\n", buf);
}
fclose(md_out);
fclose(md_of);
fclose(gt_of);
fclose(s_gt_of);
/*
* In a packed-int variant-major matrix there will be a num_vars
* number of rows, and a num_inds number of values packed into
* num_inds_ints number of intergers. For examples, 16 rows of 16 values
* will be 16 ints, where each int encodes 16 values.
*
*/
uint32_t num_var_ints = 1 + ((num_vars - 1) / 16);
uint32_t *I_data = (uint32_t *)calloc(num_var_ints*16,sizeof(uint32_t));
uint32_t **I = (uint32_t **)malloc(16*sizeof(uint32_t*));
for (i = 0; i < 16; ++i)
I[i] = I_data + i*num_var_ints;
uint32_t I_i = 0, I_int_i = 0;
uint32_t v;
s_gt_of = fopen("s.gt.tmp.packed","rb");
FILE *rs_gt_of = fopen("r.s.gt.tmp.packed","wb");
// Write these to values to that this is a well-formed uncompressed
// packed int binary file (ubin) file
fwrite(&num_vars, sizeof(uint32_t), 1, rs_gt_of);
fwrite(&num_inds, sizeof(uint32_t), 1, rs_gt_of);
/*
* we need to loop over the columns in the v-major file.
* There are num_vars rows, and num_ind_ints 16-ind packed columns
*
* In this loop :
* i: cols in var-major form, rows in ind-major form
* j: rows in var-major form, cols in ind-major form
*/
uint32_t num_inds_to_write = num_inds;
for (i = 0; i < num_ind_ints; ++i) { // loop over each int col
for (j = 0; j < num_vars; ++j) { // loop over head row in that col
// skip to the value at the row/col
fseek(s_gt_of,
j*num_ind_ints*sizeof(uint32_t) + //row
i*sizeof(uint32_t), //col
SEEK_SET);
fread(&v, sizeof(uint32_t), 1, s_gt_of);
// one int corresponds to a col of 16 two-bit values
// two_bit_i will move across the cols
for (two_bit_i = 0; two_bit_i < 16; ++two_bit_i) {
I[two_bit_i][I_i] += ((v >> (30 - 2*two_bit_i)) & 3) <<
(30 - 2*I_int_i);
}
I_int_i += 1;
if (I_int_i == 16) {
I_i += 1;
I_int_i = 0;
}
}
// When we are at the end of the file, and the number of lines
// is not a factor of 16, only write out the lines that contain values
if (num_inds_to_write >= 16) {
fwrite(I_data,
sizeof(uint32_t),
num_var_ints*16,
rs_gt_of);
num_inds_to_write -= 16;
} else {
fwrite(I_data,
sizeof(uint32_t),
num_var_ints*num_inds_to_write,
rs_gt_of);
}
memset(I_data, 0, num_var_ints*16*sizeof(uint32_t));
I_int_i = 0;
I_i = 0;
}
fclose(s_gt_of);
fclose(rs_gt_of);
free(md_index);
free(packed_ints);
}
int main_view(int argc, char *argv[])
{
int i, c, n_files = 0, out_bcf = 0, clevel = -1, multi_flag = 0, excl = 0, not_vcf = 0, in_mem = 0, u_set = 0;
long seekn = -1, n_rec = LONG_MAX, n_read = 0;
bgtm_t *bm = 0;
bcf1_t *b;
htsFile *out = 0;
char modew[8], *reg = 0, *site_flt = 0;
void *bed = 0;
int n_groups = 0;
char *gexpr[BGT_MAX_GROUPS], *aexpr = 0, *dbfn = 0, *fmt = 0;
bgt_file_t **files = 0;
fmf_t *vardb = 0;
while ((c = getopt(argc, argv, "ubs:r:l:CMGB:ef:g:a:i:n:SHt:d:")) >= 0) {
if (c == 'b') out_bcf = 1;
else if (c == 'r') reg = optarg;
else if (c == 'l') clevel = atoi(optarg);
else if (c == 'e') excl = 1;
else if (c == 'u') u_set = 1;
else if (c == 'B') bed = bed_read(optarg);
else if (c == 'C') multi_flag |= BGT_F_SET_AC;
else if (c == 'G') multi_flag |= BGT_F_NO_GT;
else if (c == 'S') multi_flag |= BGT_F_NO_GT | BGT_F_CNT_AL, not_vcf = 1;
else if (c == 'H') multi_flag |= BGT_F_NO_GT | BGT_F_CNT_HAP, not_vcf = 1;
else if (c == 'M') in_mem = 1;
else if (c == 'i') seekn = atol(optarg) - 1;
else if (c == 'n') n_rec = atol(optarg);
else if (c == 'f') site_flt = optarg;
else if (c == 't') fmt = optarg, not_vcf = 1;
else if (c == 'd') dbfn = optarg;
else if (c == 's' && n_groups < BGT_MAX_GROUPS) gexpr[n_groups++] = optarg;
else if (c == 'a') aexpr = optarg;
}
if (n_rec < 0) {
fprintf(stderr, "[E::%s] option -n must be at least 0.\n", __func__);
return 1;
}
if (clevel > 9) clevel = 9;
if (u_set) clevel = 0, out_bcf = 1;
if (n_groups > 1) multi_flag |= BGT_F_SET_AC;
if (argc - optind < 1) {
fprintf(stderr, "Usage: bgt %s [options] <bgt-prefix> [...]", argv[0]);
fputc('\n', stderr);
fprintf(stderr, "Options:\n");
fprintf(stderr, " Sample selection:\n");
fprintf(stderr, " -s EXPR samples list (,sample1,sample2 or a file or expr; see Notes below) [all]\n");
fprintf(stderr, " Site selection:\n");
fprintf(stderr, " -r STR region [all]\n");
fprintf(stderr, " -B FILE extract variants overlapping BED FILE []\n");
fprintf(stderr, " -e exclude variants overlapping BED FILE (effective with -B)\n");
fprintf(stderr, " -i INT process from the INT-th record (1-based) []\n");
fprintf(stderr, " -n INT process at most INT records []\n");
fprintf(stderr, " -d FILE variant annotations in FMF (to work with -a) []\n");
fprintf(stderr, " -M load variant annotations in RAM (only with -d)\n");
fprintf(stderr, " -a EXPR alleles list chr:1basedPos:refLen:seq (,allele1,allele2 or a file or expr) []\n");
fprintf(stderr, " -f STR frequency filters []\n");
fprintf(stderr, " VCF output:\n");
fprintf(stderr, " -b BCF output (effective without -S/-H)\n");
fprintf(stderr, " -l INT compression level for BCF [default]\n");
fprintf(stderr, " -u equivalent to -bl0 (overriding -b and -l)\n");
fprintf(stderr, " -G don't output sample genotypes\n");
fprintf(stderr, " -C write AC/AN to the INFO field (auto applied with -f or multipl -s)\n");
fprintf(stderr, " Non-VCF output:\n");
fprintf(stderr, " -S show samples with a set of alleles (with -a)\n");
fprintf(stderr, " -H count of haplotypes with a set of alleles (with -a)\n");
fprintf(stderr, " -t STR comma-delimited list of fields to output. Accepted variables:\n");
fprintf(stderr, " AC, AN, AC#, AN#, CHROM, POS, END, REF, ALT (# for a group number)\n");
fprintf(stderr, "Notes:\n");
fprintf(stderr, " For option -s/-a, EXPR can be one of:\n");
fprintf(stderr, " 1) comma-delimited list following a colon/comma. e.g. -s,NA12878,NA12044\n");
fprintf(stderr, " 2) space-delimited file with the first column giving a sample/allele name. e.g. -s list.txt\n");
fprintf(stderr, " 3) expression if .spl/-d file contains metadata. e.g.: -s\"gender=='M'&&population!='CEU'\"\n");
fprintf(stderr, " If multiple -s is specified, the AC/AN of the first group will be written to VCF INFO AC1/AN1,\n");
fprintf(stderr, " the second to AC2/AN2, etc.\n");
return 1;
}
if (dbfn && in_mem) vardb = fmf_read(dbfn), dbfn = 0;
if ((multi_flag&(BGT_F_CNT_AL|BGT_F_CNT_HAP)) && aexpr == 0) {
fprintf(stderr, "[E::%s] -a must be specified when -S/-H is in use.\n", __func__);
return 1;
}
n_files = argc - optind;
files = (bgt_file_t**)calloc(n_files, sizeof(bgt_file_t*));
for (i = 0; i < n_files; ++i) {
files[i] = bgt_open(argv[optind+i]);
if (files[i] == 0) {
fprintf(stderr, "[E::%s] failed to open BGT with prefix '%s'\n", __func__, argv[optind+i]);
return 1; // FIXME: memory leak
}
}
bm = bgtm_reader_init(n_files, files);
bgtm_set_flag(bm, multi_flag);
if (site_flt && bgtm_set_flt_site(bm, site_flt) != 0) {
fprintf(stderr, "[E::%s] failed to set frequency filters. Syntax error?\n", __func__);
return 1;
}
if (reg && bgtm_set_region(bm, reg) < 0) {
fprintf(stderr, "[E::%s] failed to set region. Region format error?\n", __func__);
return 1;
}
if (bed) bgtm_set_bed(bm, bed, excl);
if (fmt && bgtm_set_table(bm, fmt) < 0) {
fprintf(stderr, "[E::%s] failed to set tabular output.\n", __func__);
return 1;
}
if (seekn > 0) bgtm_set_start(bm, seekn);
if (aexpr) {
int n_al;
n_al = bgtm_set_alleles(bm, aexpr, vardb, dbfn);
if (n_al < 0) {
fprintf(stderr, "[E::%s] failed to set alleles.\n", __func__);
return 1;
} else if (n_al == 0)
fprintf(stderr, "[W::%s] no alleles selected.\n", __func__);
}
for (i = 0; i < n_groups; ++i) {
if (bgtm_add_group(bm, gexpr[i]) < 0) {
fprintf(stderr, "[E::%s] failed to add sample group '%s'.\n", __func__, gexpr[i]);
return 1;
}
}
bgtm_prepare(bm); // bgtm_prepare() generates the VCF header
if (!not_vcf) {
strcpy(modew, "w");
if (out_bcf) strcat(modew, "b");
sprintf(modew + strlen(modew), "%d", clevel);
out = hts_open("-", modew, 0);
vcf_hdr_write(out, bm->h_out);
}
b = bcf_init1();
while (bgtm_read(bm, b) >= 0 && n_read < n_rec) {
if (out) vcf_write1(out, bm->h_out, b);
if (fmt && bm->n_fields > 0) puts(bm->tbl_line.s);
++n_read;
}
bcf_destroy1(b);
if (not_vcf && bm->n_aal > 0) {
if (bm->flag & BGT_F_CNT_HAP) {
bgt_hapcnt_t *hc;
int n_hap;
char *s;
hc = bgtm_hapcnt(bm, &n_hap);
s = bgtm_hapcnt_print_destroy(bm, n_hap, hc);
fputs(s, stdout);
free(s);
}
if (bm->flag & BGT_F_CNT_AL) {
char *s;
if ((s = bgtm_alcnt_print(bm)) != 0)
fputs(s, stdout);
free(s);
}
}
if (out) hts_close(out);
bgtm_reader_destroy(bm);
if (bed) bed_destroy(bed);
for (i = 0; i < n_files; ++i) bgt_close(files[i]);
free(files);
if (vardb) fmf_destroy(vardb);
return 0;
}
static int mpileup(mplp_conf_t *conf)
{
if (conf->nfiles == 0) {
fprintf(stderr,"[%s] no input file/data given\n", __func__);
exit(EXIT_FAILURE);
}
mplp_ref_t mp_ref = MPLP_REF_INIT;
conf->gplp = (mplp_pileup_t *) calloc(1,sizeof(mplp_pileup_t));
conf->mplp_data = (mplp_aux_t**) calloc(conf->nfiles, sizeof(mplp_aux_t*));
conf->plp = (const bam_pileup1_t**) calloc(conf->nfiles, sizeof(bam_pileup1_t*));
conf->n_plp = (int*) calloc(conf->nfiles, sizeof(int));
// Allow to run mpileup on multiple regions in one go. This comes at cost: the bai index
// must be kept in the memory for the whole time which can be a problem with many bams.
// Therefore if none or only one region is requested, we initialize the bam iterator as
// before and free the index. Only when multiple regions are queried, we keep the index.
int nregs = 0;
if ( conf->reg_fname )
{
if ( conf->reg_is_file )
{
conf->reg = regidx_init(conf->reg_fname,NULL,NULL,0,NULL);
if ( !conf->reg ) {
fprintf(stderr,"Could not parse the regions: %s\n", conf->reg_fname);
exit(EXIT_FAILURE);
}
}
else
{
conf->reg = regidx_init(NULL,regidx_parse_reg,NULL,sizeof(char*),NULL);
if ( regidx_insert_list(conf->reg,conf->reg_fname,',') !=0 ) {
fprintf(stderr,"Could not parse the regions: %s\n", conf->reg_fname);
exit(EXIT_FAILURE);
}
}
nregs = regidx_nregs(conf->reg);
conf->reg_itr = regitr_init(conf->reg);
regitr_loop(conf->reg_itr); // region iterator now positioned at the first region
}
// read the header of each file in the list and initialize data
// beware: mpileup has always assumed that tid's are consistent in the headers, add sanity check at least!
bam_hdr_t *hdr = NULL; // header of first file in input list
int i;
for (i = 0; i < conf->nfiles; ++i) {
bam_hdr_t *h_tmp;
conf->mplp_data[i] = (mplp_aux_t*) calloc(1, sizeof(mplp_aux_t));
conf->mplp_data[i]->fp = sam_open(conf->files[i], "rb");
if ( !conf->mplp_data[i]->fp )
{
fprintf(stderr, "[%s] failed to open %s: %s\n", __func__, conf->files[i], strerror(errno));
exit(EXIT_FAILURE);
}
if (hts_set_opt(conf->mplp_data[i]->fp, CRAM_OPT_DECODE_MD, 0)) {
fprintf(stderr, "Failed to set CRAM_OPT_DECODE_MD value\n");
exit(EXIT_FAILURE);
}
if (conf->fai_fname && hts_set_fai_filename(conf->mplp_data[i]->fp, conf->fai_fname) != 0) {
fprintf(stderr, "[%s] failed to process %s: %s\n",
__func__, conf->fai_fname, strerror(errno));
exit(EXIT_FAILURE);
}
conf->mplp_data[i]->conf = conf;
conf->mplp_data[i]->ref = &mp_ref;
h_tmp = sam_hdr_read(conf->mplp_data[i]->fp);
if ( !h_tmp ) {
fprintf(stderr,"[%s] fail to read the header of %s\n", __func__, conf->files[i]);
exit(EXIT_FAILURE);
}
conf->mplp_data[i]->h = i ? hdr : h_tmp; // for j==0, "h" has not been set yet
conf->mplp_data[i]->bam_id = bam_smpl_add_bam(conf->bsmpl,h_tmp->text,conf->files[i]);
if ( conf->mplp_data[i]->bam_id<0 )
{
// no usable readgroups in this bam, it can be skipped
sam_close(conf->mplp_data[i]->fp);
free(conf->mplp_data[i]);
bam_hdr_destroy(h_tmp);
free(conf->files[i]);
if ( i+1<conf->nfiles ) memmove(&conf->files[i],&conf->files[i+1],sizeof(*conf->files)*(conf->nfiles-i-1));
conf->nfiles--;
i--;
continue;
}
if (conf->reg) {
hts_idx_t *idx = sam_index_load(conf->mplp_data[i]->fp, conf->files[i]);
if (idx == NULL) {
fprintf(stderr, "[%s] fail to load index for %s\n", __func__, conf->files[i]);
exit(EXIT_FAILURE);
}
conf->buf.l = 0;
ksprintf(&conf->buf,"%s:%u-%u",conf->reg_itr->seq,conf->reg_itr->beg+1,conf->reg_itr->end+1);
conf->mplp_data[i]->iter = sam_itr_querys(idx, conf->mplp_data[i]->h, conf->buf.s);
if ( !conf->mplp_data[i]->iter )
{
conf->mplp_data[i]->iter = sam_itr_querys(idx, conf->mplp_data[i]->h, conf->reg_itr->seq);
if ( conf->mplp_data[i]->iter ) {
fprintf(stderr,"[E::%s] fail to parse region '%s'\n", __func__, conf->buf.s);
exit(EXIT_FAILURE);
}
fprintf(stderr,"[E::%s] the sequence \"%s\" not found: %s\n",__func__,conf->reg_itr->seq,conf->files[i]);
exit(EXIT_FAILURE);
}
if ( nregs==1 ) // no need to keep the index in memory
hts_idx_destroy(idx);
else
conf->mplp_data[i]->idx = idx;
}
if ( !hdr ) hdr = h_tmp; /* save the header of first file in list */
else {
// FIXME: check consistency between h and h_tmp
bam_hdr_destroy(h_tmp);
// we store only the first file's header; it's (alleged to be)
// compatible with the i-th file's target_name lookup needs
conf->mplp_data[i]->h = hdr;
}
}
// allocate data storage proportionate to number of samples being studied sm->n
bam_smpl_get_samples(conf->bsmpl, &conf->gplp->n);
conf->gplp->n_plp = (int*) calloc(conf->gplp->n, sizeof(int));
conf->gplp->m_plp = (int*) calloc(conf->gplp->n, sizeof(int));
conf->gplp->plp = (bam_pileup1_t**) calloc(conf->gplp->n, sizeof(bam_pileup1_t*));
fprintf(stderr, "[%s] %d samples in %d input files\n", __func__, conf->gplp->n, conf->nfiles);
// write the VCF header
conf->bcf_fp = hts_open(conf->output_fname?conf->output_fname:"-", hts_bcf_wmode(conf->output_type));
if (conf->bcf_fp == NULL) {
fprintf(stderr, "[%s] failed to write to %s: %s\n", __func__, conf->output_fname? conf->output_fname : "standard output", strerror(errno));
exit(EXIT_FAILURE);
}
if ( conf->n_threads ) hts_set_threads(conf->bcf_fp, conf->n_threads);
// BCF header creation
conf->bcf_hdr = bcf_hdr_init("w");
conf->buf.l = 0;
if (conf->record_cmd_line)
{
ksprintf(&conf->buf, "##bcftoolsVersion=%s+htslib-%s\n",bcftools_version(),hts_version());
bcf_hdr_append(conf->bcf_hdr, conf->buf.s);
conf->buf.l = 0;
ksprintf(&conf->buf, "##bcftoolsCommand=mpileup");
for (i=1; i<conf->argc; i++) ksprintf(&conf->buf, " %s", conf->argv[i]);
kputc('\n', &conf->buf);
bcf_hdr_append(conf->bcf_hdr, conf->buf.s);
}
if (conf->fai_fname)
{
conf->buf.l = 0;
ksprintf(&conf->buf, "##reference=file://%s\n", conf->fai_fname);
bcf_hdr_append(conf->bcf_hdr, conf->buf.s);
}
// Translate BAM @SQ tags to BCF ##contig tags
// todo: use/write new BAM header manipulation routines, fill also UR, M5
for (i=0; i<hdr->n_targets; i++)
{
conf->buf.l = 0;
ksprintf(&conf->buf, "##contig=<ID=%s,length=%d>", hdr->target_name[i], hdr->target_len[i]);
bcf_hdr_append(conf->bcf_hdr, conf->buf.s);
}
conf->buf.l = 0;
bcf_hdr_append(conf->bcf_hdr,"##ALT=<ID=*,Description=\"Represents allele(s) other than observed.\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=INDEL,Number=0,Type=Flag,Description=\"Indicates that the variant is an INDEL.\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=IDV,Number=1,Type=Integer,Description=\"Maximum number of reads supporting an indel\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=IMF,Number=1,Type=Float,Description=\"Maximum fraction of reads supporting an indel\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=DP,Number=1,Type=Integer,Description=\"Raw read depth\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=VDB,Number=1,Type=Float,Description=\"Variant Distance Bias for filtering splice-site artefacts in RNA-seq data (bigger is better)\",Version=\"3\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=RPB,Number=1,Type=Float,Description=\"Mann-Whitney U test of Read Position Bias (bigger is better)\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=MQB,Number=1,Type=Float,Description=\"Mann-Whitney U test of Mapping Quality Bias (bigger is better)\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=BQB,Number=1,Type=Float,Description=\"Mann-Whitney U test of Base Quality Bias (bigger is better)\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=MQSB,Number=1,Type=Float,Description=\"Mann-Whitney U test of Mapping Quality vs Strand Bias (bigger is better)\">");
#if CDF_MWU_TESTS
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=RPB2,Number=1,Type=Float,Description=\"Mann-Whitney U test of Read Position Bias [CDF] (bigger is better)\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=MQB2,Number=1,Type=Float,Description=\"Mann-Whitney U test of Mapping Quality Bias [CDF] (bigger is better)\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=BQB2,Number=1,Type=Float,Description=\"Mann-Whitney U test of Base Quality Bias [CDF] (bigger is better)\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=MQSB2,Number=1,Type=Float,Description=\"Mann-Whitney U test of Mapping Quality vs Strand Bias [CDF] (bigger is better)\">");
#endif
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=SGB,Number=1,Type=Float,Description=\"Segregation based metric.\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=MQ0F,Number=1,Type=Float,Description=\"Fraction of MQ0 reads (smaller is better)\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=I16,Number=16,Type=Float,Description=\"Auxiliary tag used for calling, see description of bcf_callret1_t in bam2bcf.h\">");
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=QS,Number=R,Type=Float,Description=\"Auxiliary tag used for calling\">");
bcf_hdr_append(conf->bcf_hdr,"##FORMAT=<ID=PL,Number=G,Type=Integer,Description=\"List of Phred-scaled genotype likelihoods\">");
if ( conf->fmt_flag&B2B_FMT_DP )
bcf_hdr_append(conf->bcf_hdr,"##FORMAT=<ID=DP,Number=1,Type=Integer,Description=\"Number of high-quality bases\">");
if ( conf->fmt_flag&B2B_FMT_DV )
bcf_hdr_append(conf->bcf_hdr,"##FORMAT=<ID=DV,Number=1,Type=Integer,Description=\"Number of high-quality non-reference bases\">");
if ( conf->fmt_flag&B2B_FMT_DPR )
bcf_hdr_append(conf->bcf_hdr,"##FORMAT=<ID=DPR,Number=R,Type=Integer,Description=\"Number of high-quality bases observed for each allele\">");
if ( conf->fmt_flag&B2B_INFO_DPR )
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=DPR,Number=R,Type=Integer,Description=\"Number of high-quality bases observed for each allele\">");
if ( conf->fmt_flag&B2B_FMT_DP4 )
bcf_hdr_append(conf->bcf_hdr,"##FORMAT=<ID=DP4,Number=4,Type=Integer,Description=\"Number of high-quality ref-fwd, ref-reverse, alt-fwd and alt-reverse bases\">");
if ( conf->fmt_flag&B2B_FMT_SP )
bcf_hdr_append(conf->bcf_hdr,"##FORMAT=<ID=SP,Number=1,Type=Integer,Description=\"Phred-scaled strand bias P-value\">");
if ( conf->fmt_flag&B2B_FMT_AD )
bcf_hdr_append(conf->bcf_hdr,"##FORMAT=<ID=AD,Number=R,Type=Integer,Description=\"Allelic depths\">");
if ( conf->fmt_flag&B2B_FMT_ADF )
bcf_hdr_append(conf->bcf_hdr,"##FORMAT=<ID=ADF,Number=R,Type=Integer,Description=\"Allelic depths on the forward strand\">");
if ( conf->fmt_flag&B2B_FMT_ADR )
bcf_hdr_append(conf->bcf_hdr,"##FORMAT=<ID=ADR,Number=R,Type=Integer,Description=\"Allelic depths on the reverse strand\">");
if ( conf->fmt_flag&B2B_INFO_AD )
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=AD,Number=R,Type=Integer,Description=\"Total allelic depths\">");
if ( conf->fmt_flag&B2B_INFO_ADF )
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=ADF,Number=R,Type=Integer,Description=\"Total allelic depths on the forward strand\">");
if ( conf->fmt_flag&B2B_INFO_ADR )
bcf_hdr_append(conf->bcf_hdr,"##INFO=<ID=ADR,Number=R,Type=Integer,Description=\"Total allelic depths on the reverse strand\">");
if ( conf->gvcf )
gvcf_update_header(conf->gvcf, conf->bcf_hdr);
int nsmpl;
const char **smpl = bam_smpl_get_samples(conf->bsmpl, &nsmpl);
for (i=0; i<nsmpl; i++)
bcf_hdr_add_sample(conf->bcf_hdr, smpl[i]);
bcf_hdr_write(conf->bcf_fp, conf->bcf_hdr);
conf->bca = bcf_call_init(-1., conf->min_baseQ);
conf->bcr = (bcf_callret1_t*) calloc(nsmpl, sizeof(bcf_callret1_t));
conf->bca->openQ = conf->openQ, conf->bca->extQ = conf->extQ, conf->bca->tandemQ = conf->tandemQ;
conf->bca->min_frac = conf->min_frac;
conf->bca->min_support = conf->min_support;
conf->bca->per_sample_flt = conf->flag & MPLP_PER_SAMPLE;
conf->bc.bcf_hdr = conf->bcf_hdr;
conf->bc.n = nsmpl;
conf->bc.PL = (int32_t*) malloc(15 * nsmpl * sizeof(*conf->bc.PL));
if (conf->fmt_flag)
{
assert( sizeof(float)==sizeof(int32_t) );
conf->bc.DP4 = (int32_t*) malloc(nsmpl * sizeof(int32_t) * 4);
conf->bc.fmt_arr = (uint8_t*) malloc(nsmpl * sizeof(float)); // all fmt_flag fields, float and int32
if ( conf->fmt_flag&(B2B_INFO_DPR|B2B_FMT_DPR|B2B_INFO_AD|B2B_INFO_ADF|B2B_INFO_ADR|B2B_FMT_AD|B2B_FMT_ADF|B2B_FMT_ADR) )
{
// first B2B_MAX_ALLELES fields for total numbers, the rest per-sample
conf->bc.ADR = (int32_t*) malloc((nsmpl+1)*B2B_MAX_ALLELES*sizeof(int32_t));
conf->bc.ADF = (int32_t*) malloc((nsmpl+1)*B2B_MAX_ALLELES*sizeof(int32_t));
for (i=0; i<nsmpl; i++)
{
conf->bcr[i].ADR = conf->bc.ADR + (i+1)*B2B_MAX_ALLELES;
conf->bcr[i].ADF = conf->bc.ADF + (i+1)*B2B_MAX_ALLELES;
}
}
}
// init mpileup
conf->iter = bam_mplp_init(conf->nfiles, mplp_func, (void**)conf->mplp_data);
if ( conf->flag & MPLP_SMART_OVERLAPS ) bam_mplp_init_overlaps(conf->iter);
if ( (double)conf->max_depth * conf->nfiles > 1<<20)
fprintf(stderr, "Warning: Potential memory hog, up to %.0fM reads in the pileup!\n", (double)conf->max_depth*conf->nfiles);
if ( (double)conf->max_depth * conf->nfiles / nsmpl < 250 )
fprintf(stderr, "Note: The maximum per-sample depth with -d %d is %.1fx\n", conf->max_depth,(double)conf->max_depth * conf->nfiles / nsmpl);
bam_mplp_set_maxcnt(conf->iter, conf->max_depth);
conf->max_indel_depth = conf->max_indel_depth * nsmpl;
conf->bcf_rec = bcf_init1();
bam_mplp_constructor(conf->iter, pileup_constructor);
// Run mpileup for multiple regions
if ( nregs )
{
int ireg = 0;
do
{
// first region is already positioned
if ( ireg++ > 0 )
{
conf->buf.l = 0;
ksprintf(&conf->buf,"%s:%u-%u",conf->reg_itr->seq,conf->reg_itr->beg,conf->reg_itr->end);
for (i=0; i<conf->nfiles; i++)
{
hts_itr_destroy(conf->mplp_data[i]->iter);
conf->mplp_data[i]->iter = sam_itr_querys(conf->mplp_data[i]->idx, conf->mplp_data[i]->h, conf->buf.s);
if ( !conf->mplp_data[i]->iter )
{
conf->mplp_data[i]->iter = sam_itr_querys(conf->mplp_data[i]->idx, conf->mplp_data[i]->h, conf->reg_itr->seq);
if ( conf->mplp_data[i]->iter ) {
fprintf(stderr,"[E::%s] fail to parse region '%s'\n", __func__, conf->buf.s);
exit(EXIT_FAILURE);
}
fprintf(stderr,"[E::%s] the sequence \"%s\" not found: %s\n",__func__,conf->reg_itr->seq,conf->files[i]);
exit(EXIT_FAILURE);
}
bam_mplp_reset(conf->iter);
}
}
mpileup_reg(conf,conf->reg_itr->beg,conf->reg_itr->end);
}
while ( regitr_loop(conf->reg_itr) );
}
else
mpileup_reg(conf,0,0);
flush_bcf_records(conf, conf->bcf_fp, conf->bcf_hdr, NULL);
// clean up
free(conf->bc.tmp.s);
bcf_destroy1(conf->bcf_rec);
if (conf->bcf_fp)
{
hts_close(conf->bcf_fp);
bcf_hdr_destroy(conf->bcf_hdr);
bcf_call_destroy(conf->bca);
free(conf->bc.PL);
free(conf->bc.DP4);
free(conf->bc.ADR);
free(conf->bc.ADF);
free(conf->bc.fmt_arr);
free(conf->bcr);
}
if ( conf->gvcf ) gvcf_destroy(conf->gvcf);
free(conf->buf.s);
for (i = 0; i < conf->gplp->n; ++i) free(conf->gplp->plp[i]);
free(conf->gplp->plp); free(conf->gplp->n_plp); free(conf->gplp->m_plp); free(conf->gplp);
bam_mplp_destroy(conf->iter);
bam_hdr_destroy(hdr);
for (i = 0; i < conf->nfiles; ++i) {
if ( nregs>1 ) hts_idx_destroy(conf->mplp_data[i]->idx);
sam_close(conf->mplp_data[i]->fp);
if ( conf->mplp_data[i]->iter) hts_itr_destroy(conf->mplp_data[i]->iter);
free(conf->mplp_data[i]);
}
if ( conf->reg_itr ) regitr_destroy(conf->reg_itr);
free(conf->mplp_data); free(conf->plp); free(conf->n_plp);
free(mp_ref.ref[0]);
free(mp_ref.ref[1]);
return 0;
}
int vcf_index_stats(char *fname, int stats)
{
char *fn_out = NULL;
FILE *out;
out = fn_out ? fopen(fn_out, "w") : stdout;
const char **seq;
int i, nseq;
tbx_t *tbx = NULL;
hts_idx_t *idx = NULL;
htsFile *fp = hts_open(fname,"r");
if ( !fp ) { fprintf(stderr,"Could not read %s\n", fname); return 1; }
bcf_hdr_t *hdr = bcf_hdr_read(fp);
if ( !hdr ) { fprintf(stderr,"Could not read the header: %s\n", fname); return 1; }
if ( hts_get_format(fp)->format==vcf )
{
tbx = tbx_index_load(fname);
if ( !tbx ) { fprintf(stderr,"Could not load TBI index: %s\n", fname); return 1; }
}
else if ( hts_get_format(fp)->format==bcf )
{
idx = bcf_index_load(fname);
if ( !idx ) { fprintf(stderr,"Could not load CSI index: %s\n", fname); return 1; }
}
else
{
fprintf(stderr,"Could not detect the file type as VCF or BCF: %s\n", fname);
return 1;
}
seq = tbx ? tbx_seqnames(tbx, &nseq) : bcf_index_seqnames(idx, hdr, &nseq);
uint64_t sum = 0;
for (i=0; i<nseq; i++)
{
uint64_t records, v;
hts_idx_get_stat(tbx ? tbx->idx : idx, i, &records, &v);
sum+=records;
if (stats&2 || !records) continue;
bcf_hrec_t *hrec = bcf_hdr_get_hrec(hdr, BCF_HL_CTG, "ID", seq[i], NULL);
int hkey = hrec ? bcf_hrec_find_key(hrec, "length") : -1;
fprintf(out,"%s\t%s\t%" PRIu64 "\n", seq[i], hkey<0?".":hrec->vals[hkey], records);
}
if (!sum)
{
// No counts found.
// Is this because index version has no stored count data, or no records?
bcf1_t *rec = bcf_init1();
if (bcf_read1(fp, hdr, rec) >= 0)
{
fprintf(stderr,"%s index of %s does not contain any count metadata. Please re-index with a newer version of bcftools or tabix.\n", tbx ? "TBI" : "CSI", fname);
return 1;
}
bcf_destroy1(rec);
}
if (stats&2) fprintf(out, "%" PRIu64 "\n", sum);
free(seq);
fclose(out);
hts_close(fp);
bcf_hdr_destroy(hdr);
if (tbx)
tbx_destroy(tbx);
if (idx)
hts_idx_destroy(idx);
return 0;
}
//{{{ void print_query_result_offset(uint32_t *mask,
void print_query_result_offset(uint32_t *mask,
uint32_t mask_len,
uint32_t *vids,
struct gqt_query *q,
uint32_t **counts,
uint32_t *id_lens,
uint32_t *U_R,
uint32_t U_R_len,
char **id_query_list,
char **gt_query_list,
uint32_t num_qs,
uint32_t num_fields,
char *off_file_name,
char *source_file,
char *full_cmd)
{
struct off_file *off_f = open_off_file(off_file_name);
struct bcf_file bcf_f = init_bcf_file(source_file);
char *sample_names = NULL;
uint32_t i,j,k,line_idx,bytes, bit_i = 0;
int r;
for (i = 0; i < U_R_len; ++i) {
if (i == 0 )
r = asprintf(&sample_names,
"%s",
bcf_f.hdr->samples[U_R[i]]);
else
r = asprintf(&sample_names,
"%s,%s",
sample_names,
bcf_f.hdr->samples[U_R[i]]);
if (r == -1)
err(EX_OSERR, "asprintf error");
}
if (bcf_hdr_set_samples(bcf_f.hdr, sample_names, 0) != 0)
errx(EX_DATAERR, "Error setting samples: %s\n", source_file);
char *info_s;
for (i = 0; i < num_qs; i++) {
if ( q[i].variant_op == p_count ) {
r = asprintf(&info_s, "##INFO=<ID=GQT_%u,Number=1,Type=Integer,"
"Description=\"GQT count result from "
"phenotype:'%s' genotype:'%s'\">",
i, id_query_list[i], gt_query_list[i]);
if (r == -1) err(EX_OSERR, "asprintf error");
if (bcf_hdr_append(bcf_f.hdr, info_s) != 0)
errx(EX_DATAERR, "Error updating header: %s\n", source_file);
} else if ( q[i].variant_op == p_pct ) {
r = asprintf(&info_s, "##INFO=<ID=GQT_%u,Number=1,Type=Float,"
"Description=\"GQT percent result from "
"phenotype:'%s' genotype:'%s'\">",
i, id_query_list[i], gt_query_list[i]);
if (r == -1) err(EX_OSERR, "asprintf error");
if (bcf_hdr_append(bcf_f.hdr, info_s) != 0)
errx(EX_DATAERR, "Error updating header: %s\n", source_file);
} else if ( q[i].variant_op == p_maf ) {
r = asprintf(&info_s, "##INFO=<ID=GQT_%u,Number=1,Type=Float,"
"Description=\"GQT maf result from "
"phenotype:'%s' genotype:'%s'\">",
i, id_query_list[i], gt_query_list[i]);
if (bcf_hdr_append(bcf_f.hdr, info_s) != 0)
errx(EX_DATAERR, "Error updating header: %s\n", source_file);
}
}
r = asprintf(&info_s, "##%s_queryVersion=%s", PROGRAM_NAME, VERSION);
if (r == -1) err(EX_OSERR, "asprintf error");
if (bcf_hdr_append(bcf_f.hdr, info_s) != 0)
errx(EX_DATAERR, "Error updating header: %s\n", source_file);
r = asprintf(&info_s, "##%s_queryCommand=%s", PROGRAM_NAME, full_cmd);
if (r == -1) err(EX_OSERR, "asprintf error");
if (bcf_hdr_append(bcf_f.hdr, info_s) != 0)
errx(EX_DATAERR, "Error updating header: %s\n", source_file);
htsFile *out_f = hts_open("-","w");
if ( !out_f )
err(EX_DATAERR, "Could open output file");
bcf_hdr_write(out_f, bcf_f.hdr);
bcf_f.line = bcf_init1();
for (i=0; i < mask_len; ++i) {
bytes = mask[i];
if (bytes == 0)
continue; /* skip a bunch of ops if you can */
for (j=0; j < 32; j++) {
if (bytes & 1 << (31 - j)) {
line_idx = i*32+j;
r = goto_bcf_line(&bcf_f, off_f, line_idx);
if (r == -1)
err(EX_NOINPUT,
"Error seeking file '%s'", bcf_f.file_name);
r = get_bcf_line(&bcf_f);
if (r == -1)
err(EX_NOINPUT,
"Error reading file '%s'", bcf_f.file_name);
for (k=0; k < num_qs; k++) {
r = asprintf(&info_s, "GQT_%u", k);
if (r == -1)
err(EX_OSERR, "asprintf error");
if ( q[k].variant_op == p_count ) {
int32_t v = counts[k][line_idx];
if (bcf_update_info_int32(bcf_f.hdr,
bcf_f.line,
info_s,
&v,
1) != 0)
errx(EX_DATAERR,
"Error adding to info field: %s\n",
bcf_f.file_name);
} else if (q[k].variant_op == p_pct) {
float v = ((float)counts[k][line_idx])/
((float) id_lens[k]);
if (bcf_update_info_float(bcf_f.hdr,
bcf_f.line,
info_s,
&v,
1) != 0)
errx(EX_DATAERR,
"Error adding to info field: %s\n",
bcf_f.file_name);
} else if (q[k].variant_op == p_maf) {
float v = ((float)counts[k][line_idx])/
(((float) id_lens[k])*2.0);
if (bcf_update_info_float(bcf_f.hdr,
bcf_f.line,
info_s,
&v,
1) != 0)
errx(EX_DATAERR,
"Error adding to info field: %s\n",
bcf_f.file_name);
}
}
bcf_write(out_f, bcf_f.hdr, bcf_f.line);
}
bit_i++;
if (bit_i == num_fields)
break;
}
if (bit_i == num_fields)
break;
}
hts_close(out_f);
destroy_off_file(off_f);
}
//{{{ void get_bcf_query_result(uint32_t *mask,
void get_bcf_query_result(uint32_t *mask,
uint32_t mask_len,
struct gqt_query *q,
char **id_query_list,
uint32_t *id_lens,
uint32_t num_qs,
uint32_t num_fields,
char *vid_file_name,
char *bcf_file_name,
int bcf_output)
{
/* The VID file contains the line numbers of the variants after they have
* been sorted. To reach back into the BCF file to print the metadata
* associated with the variants marked in the mask, we need to create a
* sorted list of line numbers we want. So first we intersect the VID file
* and the mask, then sort it.
*/
/*
FILE *vid_f = fopen(vid_file_name, "rb");
if (!vid_f)
err(EX_NOINPUT, "Cannot read file\"%s\"", vid_file_name);
uint32_t *vids = (uint32_t *) malloc(num_fields*sizeof(uint32_t));
if (!vids )
err(EX_OSERR, "malloc error");
size_t fr = fread(vids, sizeof(uint32_t), num_fields, vid_f);
check_file_read(vid_file_name, vid_f, num_fields, fr);
fclose(vid_f);
*/
struct vid_file *vid_f = open_vid_file(vid_file_name);
load_vid_data(vid_f);
uint32_t i, j, masked_vid_count = 0;
for (i = 0; i < mask_len; ++i)
masked_vid_count += popcount(mask[i]);
uint32_t *masked_vids = (uint32_t *)
malloc(masked_vid_count*sizeof(uint32_t));
if (!masked_vids )
err(EX_OSERR, "malloc error");
uint32_t masked_vid_i = 0;
for (i = 0; i < mask_len; ++i) {
uint32_t bytes = mask[i];
if (bytes == 0)
continue; /* skip a bunch of ops if you can */
for (j = 0; j < 32; j++) {
if (bytes & (1 << (31 - j))) {
masked_vids[masked_vid_i] = vid_f->vids[i*32 + j];
masked_vid_i+=1;
}
}
if (masked_vid_i == masked_vid_count)
break;
}
destroy_vid_file(vid_f);
qsort(masked_vids, masked_vid_count, sizeof(uint32_t), compare_uint32_t);
htsFile *fp = hts_open(bcf_file_name,"rb");
bcf_hdr_t *hdr = bcf_hdr_read(fp);
bcf1_t *line = bcf_init1();
//bcf_hdr_set_samples(hdr, print_name_csv, 0);
htsFile *out;
if (!bcf_output)
out = hts_open("-", "w");
else
out = hts_open("-", "wb");
int r = bcf_hdr_write(out, hdr);
uint32_t bcf_line_i = 0;
masked_vid_i = 0;
while ( bcf_read(fp, hdr, line) != -1) {
if (masked_vids[masked_vid_i] == bcf_line_i) {
r = bcf_unpack(line, BCF_UN_ALL);
r = bcf_write1(out, hdr, line);
masked_vid_i+=1;
}
if (masked_vid_i == masked_vid_count)
break;
bcf_line_i += 1;
}
hts_close(out);
hts_close(fp);
}
int main_vcfview(int argc, char *argv[])
{
int i, c, clevel = -1, flag = 0, n_samples = -1, *imap = 0, excl_snp = 0, excl_indel = 0;
char *fn_ref = 0, *fn_out = 0, moder[8], **samples = 0;
bcf_hdr_t *h, *hsub = 0;
htsFile *in;
bcf1_t *b;
while ((c = getopt(argc, argv, "l:bSt:o:T:s:GNI")) >= 0) {
switch (c) {
case 'l': clevel = atoi(optarg); flag |= 2; break;
case 'S': flag |= 1; break;
case 'b': flag |= 2; break;
case 'G': n_samples = 0; break;
case 't': fn_ref = optarg; flag |= 1; break;
case 'o': fn_out = optarg; break;
case 's': samples = hts_readlines(optarg, &n_samples); break;
case 'N': excl_snp = 1; break;
case 'I': excl_indel = 1; break;
}
}
if (argc == optind) {
fprintf(stderr, "\nUsage: vcfview [options] <in.bcf>|<in.vcf>|<in.vcf.gz>\n\n");
fprintf(stderr, "Options: -b output in BCF\n");
fprintf(stderr, " -S input is VCF\n");
fprintf(stderr, " -o FILE output file name [stdout]\n");
fprintf(stderr, " -l INT compression level [%d]\n", clevel);
fprintf(stderr, " -t FILE list of reference names and lengths [null]\n");
fprintf(stderr, " -s FILE/STR list of samples (STR if started with ':'; FILE otherwise) [null]\n");
fprintf(stderr, " -G drop individual genotype information\n");
fprintf(stderr, " -N exclude SNPs\n");
fprintf(stderr, " -I exclude INDELs\n");
fprintf(stderr, "\n");
return 1;
}
strcpy(moder, "r");
if ((flag&1) == 0 && !(file_type(argv[optind])&(IS_VCF|IS_VCF_GZ))) strcat(moder, "b");
in = hts_open(argv[optind], moder, fn_ref);
h = vcf_hdr_read(in);
if (h == 0) {
fprintf(stderr, "[E::%s] fail to read the VCF/BCF2 header\n", __func__);
hts_close(in);
return 1;
}
if (n_samples >= 0) {
if (n_samples) imap = (int*)malloc(n_samples * sizeof(int));
hsub = bcf_hdr_subset(h, n_samples, samples, imap);
}
b = bcf_init1();
if ((flag&4) == 0) { // VCF/BCF output
htsFile *out;
char modew[8];
strcpy(modew, "w");
if (clevel >= 0 && clevel <= 9) sprintf(modew + 1, "%d", clevel);
if (flag&2) strcat(modew, "b");
out = hts_open(fn_out? fn_out : "-", modew, 0);
vcf_hdr_write(out, hsub? hsub : h);
if (optind + 1 < argc && !(flag&1)) { // BAM input and has a region
hts_idx_t *idx;
if ((idx = bcf_index_load(argv[optind])) == 0) {
fprintf(stderr, "[E::%s] fail to load the BCF index\n", __func__);
return 1;
}
for (i = optind + 1; i < argc; ++i) {
hts_itr_t *iter;
if ((iter = bcf_itr_querys(idx, h, argv[i])) == 0) {
fprintf(stderr, "[E::%s] fail to parse region '%s'\n", __func__, argv[i]);
continue;
}
while (bcf_itr_next((BGZF*)in->fp, iter, b) >= 0) {
if (excl_snp && bcf_is_snp(b)) continue;
if (excl_indel && !bcf_is_snp(b)) continue;
if (n_samples >= 0) {
bcf_subset(h, b, n_samples, imap);
vcf_write1(out, hsub, b);
} else vcf_write1(out, h, b);
}
hts_itr_destroy(iter);
}
hts_idx_destroy(idx);
} else {
while (vcf_read1(in, h, b) >= 0) {
if (excl_snp && bcf_is_snp(b)) continue;
if (excl_indel && !bcf_is_snp(b)) continue;
if (n_samples >= 0) {
bcf_subset(h, b, n_samples, imap);
vcf_write1(out, hsub, b);
} else vcf_write1(out, h, b);
}
}
hts_close(out);
}
bcf_destroy1(b);
if (n_samples > 0) {
for (i = 0; i < n_samples; ++i) free(samples[i]);
free(samples);
bcf_hdr_destroy(hsub);
free(imap);
}
bcf_hdr_destroy(h);
hts_close(in);
return 0;
}
int bcf_sr_next_line(readers_t *files)
{
int32_t min_pos = INT_MAX;
int ret,i,j;
kstring_t *str = &files->tmps;
while ( min_pos==INT_MAX )
{
// Need to open new chromosome?
int eos = 0;
for (i=0; i<files->nreaders; i++)
if ( !files->readers[i].itr && !files->readers[i].nbuffer ) eos++;
if ( eos==files->nreaders )
{
const char *seq;
if ( files->targets )
{
seq = tgt_next_seq(files->targets);
if ( !seq ) return 0; // all chroms scanned
}
else
{
if ( ++files->iseq >= files->nseqs ) return 0; // all chroms scanned
seq = files->seqs[files->iseq];
}
for (i=0; i<files->nreaders; i++)
{
reader_t *reader = &files->readers[i];
if ( reader->tbx )
reader->itr = tbx_itr_querys(reader->tbx,seq);
else
reader->itr = bcf_itr_querys(reader->bcf,reader->header,seq);
}
}
// Find the smallest coordinate
for (i=0; i<files->nreaders; i++)
{
reader_t *reader = &files->readers[i];
int buffer_full = ( reader->nbuffer && reader->buffer[reader->nbuffer]->pos != reader->buffer[1]->pos ) ? 1 : 0;
if ( reader->itr && !buffer_full )
{
// Fill the buffer with records starting at the same position
while (1)
{
if ( reader->nbuffer+1 >= reader->mbuffer )
{
reader->mbuffer += 8;
reader->buffer = (bcf1_t**) realloc(reader->buffer, sizeof(bcf1_t*)*reader->mbuffer);
for (j=8; j>0; j--)
reader->buffer[reader->mbuffer-j] = bcf_init1();
}
if ( reader->tbx )
{
ret = tbx_itr_next((BGZF*)reader->file->fp, reader->tbx, reader->itr, str);
if ( ret<0 ) break;
vcf_parse1(str, reader->header, reader->buffer[reader->nbuffer+1]);
}
else
{
ret = bcf_itr_next((BGZF*)reader->file->fp, reader->itr, reader->buffer[reader->nbuffer+1]);
if ( ret<0 ) break;
}
bcf_unpack(reader->buffer[reader->nbuffer+1], BCF_UN_STR|BCF_UN_FLT);
// apply filter
if ( reader->filter_id!=-1 && reader->buffer[reader->nbuffer+1]->d.n_flt && reader->filter_id!=reader->buffer[reader->nbuffer+1]->d.flt[0] ) continue;
set_variant_types(reader->buffer[reader->nbuffer+1]);
reader->nbuffer++;
if ( reader->buffer[reader->nbuffer]->pos != reader->buffer[1]->pos ) break;
}
if ( ret<0 ) { tbx_itr_destroy(reader->itr); reader->itr = NULL; } // done for this chromosome
}
if ( reader->nbuffer )
{
if ( min_pos > reader->buffer[1]->pos ) min_pos = reader->buffer[1]->pos;
}
// The buffer is full - either there is nothing else to read or the last record has a different coordinate
if ( files->collapse && reader->nbuffer>2 && reader->buffer[1]->pos==reader->buffer[2]->pos )
{
collapse_buffer(files, reader);
}
}
if ( files->targets && min_pos!=INT_MAX )
{
int ret = tgt_has_position(files->targets, min_pos);
if ( ret==1 ) continue;
// The position must be skipped
if ( ret==-1 )
{
// done for this chromosome, don't read the rest
for (i=0; i<files->nreaders; i++)
{
files->readers[i].nbuffer = 0;
if ( files->readers[i].itr )
{
tbx_itr_destroy(files->readers[i].itr);
files->readers[i].itr = NULL;
}
}
min_pos = INT_MAX;
continue;
}
// remove the active line, save the buffer line
for (i=0; i<files->nreaders; i++)
{
reader_t *reader = &files->readers[i];
for (j=1; j<=reader->nbuffer; j++)
if ( reader->buffer[j]->pos!=min_pos ) break;
if ( j==1 ) continue;
if ( j<=reader->nbuffer )
{
bcf1_t *tmp = reader->buffer[1]; reader->buffer[1] = reader->buffer[j]; reader->buffer[j] = tmp;
reader->nbuffer = 1;
}
else
reader->nbuffer = 0;
}
min_pos = INT_MAX;
}
}
//printf("[next_line] min_pos=%d\n", min_pos+1);
//debug_buffers(files);
// Set the current line
ret = 0;
bcf1_t *first = NULL;
for (i=0; i<files->nreaders; i++)
{
reader_t *reader = &files->readers[i];
if ( !reader->nbuffer || reader->buffer[1]->pos!=min_pos ) continue;
// Match the records by REF and ALT
int j, irec = -1;
if ( first )
{
for (j=1; j<=reader->nbuffer; j++)
{
bcf1_t *line = reader->buffer[j];
if ( min_pos != line->pos ) break; // done with this buffer
if ( files->collapse&COLLAPSE_ANY ) { irec=j; break; } // checking position only
if ( files->collapse&COLLAPSE_SNPS && first->d.var_type&VCF_SNP && line->d.var_type&VCF_SNP ) { irec=j; break; }
if ( files->collapse&COLLAPSE_INDELS && first->d.var_type&VCF_INDEL && line->d.var_type&VCF_INDEL ) { irec=j; break; }
if ( first->rlen != line->rlen ) continue; // REFs do not match
if ( strcmp(first->d.allele[0], line->d.allele[0]) ) continue; // REFs do not match
int ial,jal;
if ( files->collapse==COLLAPSE_NONE )
{
// require exact match, all alleles must be identical
if ( first->n_allele!=line->n_allele ) continue; // different number of alleles
int nmatch = 1; // REF has been already checked
for (ial=1; ial<first->n_allele; ial++)
{
for (jal=1; jal<line->n_allele; jal++)
if ( !strcmp(first->d.allele[ial], line->d.allele[jal]) ) { nmatch++; break; }
}
if ( nmatch>=first->n_allele ) { irec=j; break; }
}
else
{
// thorough check: the REFs and some of the alleles have to be shared
// (neglecting different representations of the same indel for now)
for (ial=1; ial<first->n_allele; ial++)
{
for (jal=1; jal<line->n_allele; jal++)
if ( !strcmp(first->d.allele[ial], line->d.allele[jal]) ) { irec=j; break; }
if ( irec>=1 ) break;
}
}
if ( irec>=1 ) break;
}
if ( irec==-1 ) continue;
}
else
{
first = reader->buffer[1];
irec = 1;
}
bcf1_t *tmp = reader->buffer[0];
reader->buffer[0] = reader->buffer[irec];
for (j=irec+1; j<=reader->nbuffer; j++)
reader->buffer[j-1] = reader->buffer[j];
reader->buffer[ reader->nbuffer ] = tmp;
reader->nbuffer--;
ret |= 1<<i;
}
// fprintf(stdout,"[next_line] min_pos=%d mask=%d\n", min_pos+1, ret);
// debug_buffers(stdout,files);
return ret;
}
int main(int argc, char **argv)
{
char *fname = argc>1 ? argv[1] : "/dev/null";
htsFile *fp = hts_open(fname, "w");
bcf_hdr_t *hdr1, *hdr2;
hdr1 = bcf_hdr_init("w");
hdr2 = bcf_hdr_init("w");
// Add two shared and two private annotations
bcf_hdr_append(hdr1, "##contig=<ID=1>");
bcf_hdr_append(hdr1, "##contig=<ID=2>");
bcf_hdr_append(hdr2, "##contig=<ID=2>");
bcf_hdr_append(hdr2, "##contig=<ID=1>");
bcf_hdr_append(hdr1, "##FILTER=<ID=FLT1,Description=\"Filter 1\">");
bcf_hdr_append(hdr1, "##FILTER=<ID=FLT2,Description=\"Filter 2\">");
bcf_hdr_append(hdr1, "##FILTER=<ID=FLT3,Description=\"Filter 3\">");
bcf_hdr_append(hdr2, "##FILTER=<ID=FLT4,Description=\"Filter 4\">");
bcf_hdr_append(hdr2, "##FILTER=<ID=FLT3,Description=\"Filter 3\">");
bcf_hdr_append(hdr2, "##FILTER=<ID=FLT2,Description=\"Filter 2\">");
bcf_hdr_append(hdr1, "##INFO=<ID=INF1,Number=.,Type=Integer,Description=\"Info 1\">");
bcf_hdr_append(hdr1, "##INFO=<ID=INF2,Number=.,Type=Integer,Description=\"Info 2\">");
bcf_hdr_append(hdr1, "##INFO=<ID=INF3,Number=.,Type=Integer,Description=\"Info 3\">");
bcf_hdr_append(hdr2, "##INFO=<ID=INF4,Number=.,Type=Integer,Description=\"Info 4\">");
bcf_hdr_append(hdr2, "##INFO=<ID=INF3,Number=.,Type=Integer,Description=\"Info 3\">");
bcf_hdr_append(hdr2, "##INFO=<ID=INF2,Number=.,Type=Integer,Description=\"Info 2\">");
bcf_hdr_append(hdr1, "##FORMAT=<ID=FMT1,Number=.,Type=Integer,Description=\"FMT 1\">");
bcf_hdr_append(hdr1, "##FORMAT=<ID=FMT2,Number=.,Type=Integer,Description=\"FMT 2\">");
bcf_hdr_append(hdr1, "##FORMAT=<ID=FMT3,Number=.,Type=Integer,Description=\"FMT 3\">");
bcf_hdr_append(hdr2, "##FORMAT=<ID=FMT4,Number=.,Type=Integer,Description=\"FMT 4\">");
bcf_hdr_append(hdr2, "##FORMAT=<ID=FMT3,Number=.,Type=Integer,Description=\"FMT 3\">");
bcf_hdr_append(hdr2, "##FORMAT=<ID=FMT2,Number=.,Type=Integer,Description=\"FMT 2\">");
bcf_hdr_add_sample(hdr1,"SMPL1");
bcf_hdr_add_sample(hdr1,"SMPL2");
bcf_hdr_add_sample(hdr2,"SMPL1");
bcf_hdr_add_sample(hdr2,"SMPL2");
bcf_hdr_sync(hdr1);
bcf_hdr_sync(hdr2);
hdr2 = bcf_hdr_merge(hdr2,hdr1);
bcf_hdr_sync(hdr2);
if ( bcf_hdr_write(fp, hdr2)!=0 ) error("Failed to write to %s\n", fname);
bcf1_t *rec = bcf_init1();
rec->rid = bcf_hdr_name2id(hdr1, "1");
rec->pos = 0;
bcf_update_alleles_str(hdr1, rec, "G,A");
int32_t tmpi[3];
tmpi[0] = bcf_hdr_id2int(hdr1, BCF_DT_ID, "FLT1");
tmpi[1] = bcf_hdr_id2int(hdr1, BCF_DT_ID, "FLT2");
tmpi[2] = bcf_hdr_id2int(hdr1, BCF_DT_ID, "FLT3");
bcf_update_filter(hdr1, rec, tmpi, 3);
tmpi[0] = 1; bcf_update_info_int32(hdr1, rec, "INF1", tmpi, 1);
tmpi[0] = 2; bcf_update_info_int32(hdr1, rec, "INF2", tmpi, 1);
tmpi[0] = 3; bcf_update_info_int32(hdr1, rec, "INF3", tmpi, 1);
tmpi[0] = tmpi[1] = 1; bcf_update_format_int32(hdr1, rec, "FMT1", tmpi, 2);
tmpi[0] = tmpi[1] = 2; bcf_update_format_int32(hdr1, rec, "FMT2", tmpi, 2);
tmpi[0] = tmpi[1] = 3; bcf_update_format_int32(hdr1, rec, "FMT3", tmpi, 2);
bcf_remove_filter(hdr1, rec, bcf_hdr_id2int(hdr1, BCF_DT_ID, "FLT2"), 0);
bcf_update_info_int32(hdr1, rec, "INF2", NULL, 0);
bcf_update_format_int32(hdr1, rec, "FMT2", NULL, 0);
bcf_translate(hdr2, hdr1, rec);
if ( bcf_write(fp, hdr2, rec)!=0 ) error("Faild to write to %s\n", fname);
// Clean
bcf_destroy1(rec);
bcf_hdr_destroy(hdr1);
bcf_hdr_destroy(hdr2);
int ret;
if ( (ret=hts_close(fp)) )
{
fprintf(stderr,"hts_close(%s): non-zero status %d\n",fname,ret);
exit(ret);
}
return 0;
}
static void buffered_filters(args_t *args, bcf1_t *line)
{
/**
* The logic of SnpGap=3. The SNPs at positions 1 and 7 are filtered,
* positions 0 and 8 are not:
* 0123456789
* ref .G.GT..G..
* del .A.G-..A..
* Here the positions 1 and 6 are filtered, 0 and 7 are not:
* 0123-456789
* ref .G.G-..G..
* ins .A.GT..A..
*
* The logic of IndelGap=2. The second indel is filtered:
* 012345678901
* ref .GT.GT..GT..
* del .G-.G-..G-..
* And similarly here, the second is filtered:
* 01 23 456 78
* ref .A-.A-..A-..
* ins .AT.AT..AT..
*/
// To avoid additional data structure, we abuse bcf1_t's var and var_type records.
const int SnpGap_set = VCF_OTHER<<1;
const int IndelGap_set = VCF_OTHER<<2;
const int IndelGap_flush = VCF_OTHER<<3;
int var_type = 0, i;
if ( line )
{
// Still on the same chromosome?
int ilast = rbuf_last(&args->rbuf);
if ( ilast>=0 && line->rid != args->rbuf_lines[ilast]->rid )
flush_buffer(args, args->rbuf.n); // new chromosome, flush everything
rbuf_expand0(&args->rbuf,bcf1_t*,args->rbuf.n,args->rbuf_lines);
// Insert the new record in the buffer. The line would be overwritten in
// the next bcf_sr_next_line call, therefore we need to swap it with an
// unused one
ilast = rbuf_append(&args->rbuf);
if ( !args->rbuf_lines[ilast] ) args->rbuf_lines[ilast] = bcf_init1();
SWAP(bcf1_t*, args->files->readers[0].buffer[0], args->rbuf_lines[ilast]);
var_type = bcf_get_variant_types(line);
// Find out the size of an indel. The indel boundaries are based on REF
// (POS+1,POS+rlen-1). This is not entirely correct: mpileup likes to
// output REF=CAGAGAGAGA, ALT=CAGAGAGAGAGA where REF=C,ALT=CGA could be
// used. This filter is therefore more strict and may remove some valid
// SNPs.
int len = 1;
if ( var_type & VCF_INDEL )
{
for (i=1; i<line->n_allele; i++)
if ( len < 1-line->d.var[i].n ) len = 1-line->d.var[i].n;
}
// Set the REF allele's length to max deletion length or to 1 if a SNP or an insertion.
line->d.var[0].n = len;
}
int k_flush = 1;
if ( args->indel_gap )
{
k_flush = 0;
// Find indels which are too close to each other
int last_to = -1;
for (i=-1; rbuf_next(&args->rbuf,&i); )
{
bcf1_t *rec = args->rbuf_lines[i];
int rec_from = rec->pos;
if ( last_to!=-1 && last_to < rec_from ) break;
k_flush++;
if ( !(rec->d.var_type & VCF_INDEL) ) continue;
rec->d.var_type |= IndelGap_set;
last_to = args->indel_gap + rec->pos + rec->d.var[0].n - 1;
}
if ( i==args->rbuf.f && line && last_to!=-1 ) k_flush = 0;
if ( k_flush || !line )
{
// Select the best indel from the cluster of k_flush indels
int k = 0, max_ac = -1, imax_ac = -1;
for (i=-1; rbuf_next(&args->rbuf,&i) && k<k_flush; )
{
k++;
bcf1_t *rec = args->rbuf_lines[i];
if ( !(rec->d.var_type & IndelGap_set) ) continue;
hts_expand(int, rec->n_allele, args->ntmpi, args->tmpi);
int ret = bcf_calc_ac(args->hdr, rec, args->tmpi, BCF_UN_ALL);
if ( imax_ac==-1 || (ret && max_ac < args->tmpi[1]) ) { max_ac = args->tmpi[1]; imax_ac = i; }
}
// Filter all but the best indel (with max AF or first if AF not available)
k = 0;
for (i=-1; rbuf_next(&args->rbuf,&i) && k<k_flush; )
{
k++;
bcf1_t *rec = args->rbuf_lines[i];
if ( !(rec->d.var_type & IndelGap_set) ) continue;
rec->d.var_type |= IndelGap_flush;
if ( i!=imax_ac ) bcf_add_filter(args->hdr, args->rbuf_lines[i], args->IndelGap_id);
}
}
int main(int argc, char **argv)
{
int i, n;
static struct option const long_opts[] =
{
{"out", required_argument, NULL, 1},
{"report", required_argument, NULL, 2},
{"dotasref", no_argument, NULL, 3},
{"help", no_argument, NULL, 0},
{"version", no_argument, NULL, 4},
{"export_uncov", no_argument, NULL, 5}
};
bool help = FALSE;
bool report_version = FALSE;
while ((n = getopt_long(argc, argv, "1:2:304", long_opts, NULL)) >= 0)
{
switch (n)
{
case 1 : outfile = strdup(optarg); break;
case 2 : report = strdup(optarg); break;
case 3 : dotasref = TRUE; break;
case 0 : help = TRUE; break;
case 4 : report_version = TRUE; break;
case 5 : export_uncover = TRUE; break;
default : return 1;
}
if ( help ) return usage();
if ( report_version ) return show_version();
}
n = argc - optind;
if ( n > 1 ) errabort("only accept one input vcf");
if ( export_uncover == TRUE && outfile == FALSE) {
warnings("export uncove region only used with option --out");
export_uncover = FALSE;
}
char * input;
if ( n == 0 ) input = strdup("-");
else input = strdup(argv[optind]);
htsFile * fp = read_vcf_file(input);
enum htsExactFormat fmt = hts_get_format(fp)->format;
if ( fmt != vcf && fmt != bcf ) errabort("This is not a VCF/BCF file : %s", input);
bcf_hdr_t * hdr = bcf_hdr_read(fp);
int n_samples = bcf_hdr_nsamples(hdr);
if ( n_samples != 2 ) errabort("the input VCF/BCF file must contain only two samples! %d", n_samples);
LOG("Using sample %s as ref ...", hdr->samples[0]);
LOG("Using sample %s as test ...", hdr->samples[1]);
uint32_t matrix[4][4] = { {0, 0, 0, 0}, {0, 0, 0, 0}, {0, 0, 0, 0}, {0, 0, 0, 0} };
bcf1_t * v = bcf_init1();
kstring_t str = { 0, 0, 0 };
uint32_t line = 0;
htsFile *out = NULL;
if ( outfile && !check_filename(outfile) ) out = hts_open(outfile, mode);
if ( out != NULL ) bcf_hdr_write(out, hdr);
while ( bcf_read1(fp, hdr, v) >= 0 )
{
bcf_unpack(v, BCF_UN_STR|BCF_UN_FMT);
int k;
str.l = 0;
int tag_id = bcf_hdr_id2int(hdr, BCF_DT_ID, "GT");
if ( !bcf_hdr_idinfo_exists(hdr, BCF_HL_FMT, tag_id) ) warnings("There is no 'GT' in the header!");
for ( i = 0; i < v->n_fmt; ++i )
if ( v->d.fmt[i].id == tag_id ) break;
if ( i == v->n_fmt ) {
vcf_format1(hdr, v, &str);
LOG("There is no tag GT in this line : %s", str.s);
continue;
}
corr_t xy[2] = { {-1, -2, -2}, {-1, -2, -2} };
bcf_fmt_t * fmt = &v->d.fmt[i];
for ( i = 0; i < 2; ++i )
{
int corr = i;
if ( fmt == NULL ) {
if ( dotasref == TRUE ) xy[corr].alt = ALT_IS_REF;
else xy[corr].alt = ALT_IS_UNC;
continue;
}
int last = -2;
uint8_t *d = (uint8_t*)((char*)fmt->p + fmt->size*i);
for ( k = 0; k < fmt->n && d[k] != (uint8_t)bcf_int8_vector_end; ++k )
{
int curr = d[k]>>1;
if ( last != curr ) {
if ( curr ) {
if ( last == -2 ) xy[corr].alt = curr > 1 ? ALT_IS_HOM : ALT_IS_REF;
else xy[corr].alt = ALT_IS_HET;
} else {
xy[corr].alt = dotasref == TRUE ? ALT_IS_REF : ALT_IS_UNC;
}
} else {
if ( curr ) {
xy[corr].alt = curr > 1 ? ALT_IS_HOM : ALT_IS_REF;
} else {
xy[corr].alt = dotasref == TRUE ? ALT_IS_REF : ALT_IS_UNC;
}
}
if (last == -2 ) {
xy[corr].min = xy[corr].max = curr;
} else {
if ( curr < xy[corr].min ) xy[corr].min = curr;
else if ( curr > xy[corr].max ) xy[corr].max = curr;
}
last = curr;
}
}
matrix[xy[0].alt][xy[1].alt]++;
if ( xy[0].alt != xy[1].alt && out != NULL) {
if ( xy[0].alt == ALT_IS_UNC || xy[1].alt == ALT_IS_UNC ) {
if ( export_uncover == TRUE ) {
str.l = 0;
vcf_format1(hdr, v, &str);
vcf_write(out, hdr, v);
}
} else {
str.l = 0;
vcf_format1(hdr, v, &str);
vcf_write(out, hdr, v);
}
}
if ( xy[0].alt == ALT_IS_HET && xy[1].alt == ALT_IS_HET && (xy[0].min != xy[1].min || xy[0].max != xy[1].max ) ) {
bias++;
matrix[ALT_IS_HET][ALT_IS_HET]--;
if ( out != NULL ) {
str.l = 0;
vcf_format1(hdr, v, &str);
vcf_write(out, hdr, v);
}
}
line++;
}
if ( out ) hts_close(out);
if ( str.m ) free(str.s);
write_report(matrix, hdr);
bcf_hdr_destroy(hdr);
free(input);
bcf_destroy1(v);
if ( outfile ) free(outfile);
if ( report ) free(report);
if ( hts_close(fp) ) warnings("hts_close returned non-zero status: %s", input);
return 0;
}
/*
* _reader_fill_buffer() - buffers all records with the same coordinate
*/
static void _reader_fill_buffer(bcf_srs_t *files, bcf_sr_t *reader)
{
// Return if the buffer is full: the coordinate of the last buffered record differs
if ( reader->nbuffer && reader->buffer[reader->nbuffer]->pos != reader->buffer[1]->pos ) return;
// No iterator (sequence not present in this file) and not streaming
if ( !reader->itr && !files->streaming ) return;
// Fill the buffer with records starting at the same position
int i, ret = 0;
while (1)
{
if ( reader->nbuffer+1 >= reader->mbuffer )
{
// Increase buffer size
reader->mbuffer += 8;
reader->buffer = (bcf1_t**) realloc(reader->buffer, sizeof(bcf1_t*)*reader->mbuffer);
for (i=8; i>0; i--) // initialize
{
reader->buffer[reader->mbuffer-i] = bcf_init1();
reader->buffer[reader->mbuffer-i]->max_unpack = files->max_unpack;
reader->buffer[reader->mbuffer-i]->pos = -1; // for rare cases when VCF starts from 1
}
}
if ( files->streaming )
{
if ( reader->file->format.format==vcf )
{
if ( (ret=hts_getline(reader->file, KS_SEP_LINE, &files->tmps)) < 0 ) break; // no more lines
int ret = vcf_parse1(&files->tmps, reader->header, reader->buffer[reader->nbuffer+1]);
if ( ret<0 ) break;
}
else if ( reader->file->format.format==bcf )
{
if ( (ret=bcf_read1(reader->file, reader->header, reader->buffer[reader->nbuffer+1])) < 0 ) break; // no more lines
}
else
{
fprintf(stderr,"[%s:%d %s] fixme: not ready for this\n", __FILE__,__LINE__,__FUNCTION__);
exit(1);
}
}
else if ( reader->tbx_idx )
{
if ( (ret=tbx_itr_next(reader->file, reader->tbx_idx, reader->itr, &files->tmps)) < 0 ) break; // no more lines
vcf_parse1(&files->tmps, reader->header, reader->buffer[reader->nbuffer+1]);
}
else
{
if ( (ret=bcf_itr_next(reader->file, reader->itr, reader->buffer[reader->nbuffer+1])) < 0 ) break; // no more lines
bcf_subset_format(reader->header,reader->buffer[reader->nbuffer+1]);
}
// apply filter
if ( !reader->nfilter_ids )
bcf_unpack(reader->buffer[reader->nbuffer+1], BCF_UN_STR);
else
{
bcf_unpack(reader->buffer[reader->nbuffer+1], BCF_UN_STR|BCF_UN_FLT);
if ( !has_filter(reader, reader->buffer[reader->nbuffer+1]) ) continue;
}
reader->nbuffer++;
if ( reader->buffer[reader->nbuffer]->pos != reader->buffer[1]->pos ) break; // the buffer is full
}
if ( ret<0 )
{
// done for this region
tbx_itr_destroy(reader->itr);
reader->itr = NULL;
}
if ( files->collapse && reader->nbuffer>=2 && reader->buffer[1]->pos==reader->buffer[2]->pos )
collapse_buffer(files, reader);
}
/*
* Performs pileup
* @param conf configuration for this pileup
* @param n number of files specified in fn
* @param fn filenames
*/
static int mpileup(mplp_conf_t *conf, int n, char **fn)
{
extern void *bcf_call_add_rg(void *rghash, const char *hdtext, const char *list);
extern void bcf_call_del_rghash(void *rghash);
mplp_aux_t **data;
int i, tid, pos, *n_plp, tid0 = -1, beg0 = 0, end0 = 1u<<29, ref_len, ref_tid = -1, max_depth, max_indel_depth;
const bam_pileup1_t **plp;
bam_mplp_t iter;
bam_hdr_t *h = NULL; /* header of first file in input list */
char *ref;
void *rghash = NULL;
FILE *pileup_fp = NULL;
bcf_callaux_t *bca = NULL;
bcf_callret1_t *bcr = NULL;
bcf_call_t bc;
htsFile *bcf_fp = NULL;
bcf_hdr_t *bcf_hdr = NULL;
bam_sample_t *sm = NULL;
kstring_t buf;
mplp_pileup_t gplp;
memset(&gplp, 0, sizeof(mplp_pileup_t));
memset(&buf, 0, sizeof(kstring_t));
memset(&bc, 0, sizeof(bcf_call_t));
data = calloc(n, sizeof(mplp_aux_t*));
plp = calloc(n, sizeof(bam_pileup1_t*));
n_plp = calloc(n, sizeof(int));
sm = bam_smpl_init();
if (n == 0) {
fprintf(stderr,"[%s] no input file/data given\n", __func__);
exit(1);
}
// read the header of each file in the list and initialize data
for (i = 0; i < n; ++i) {
bam_hdr_t *h_tmp;
data[i] = calloc(1, sizeof(mplp_aux_t));
data[i]->fp = sam_open(fn[i], "rb");
if ( !data[i]->fp )
{
fprintf(stderr, "[%s] failed to open %s: %s\n", __func__, fn[i], strerror(errno));
exit(1);
}
hts_set_fai_filename(data[i]->fp, conf->fai_fname);
data[i]->conf = conf;
h_tmp = sam_hdr_read(data[i]->fp);
if ( !h_tmp ) {
fprintf(stderr,"[%s] fail to read the header of %s\n", __func__, fn[i]);
exit(1);
}
data[i]->h = i? h : h_tmp; // for i==0, "h" has not been set yet
bam_smpl_add(sm, fn[i], (conf->flag&MPLP_IGNORE_RG)? 0 : h_tmp->text);
// Collect read group IDs with PL (platform) listed in pl_list (note: fragile, strstr search)
rghash = bcf_call_add_rg(rghash, h_tmp->text, conf->pl_list);
if (conf->reg) {
hts_idx_t *idx = sam_index_load(data[i]->fp, fn[i]);
if (idx == 0) {
fprintf(stderr, "[%s] fail to load index for %s\n", __func__, fn[i]);
exit(1);
}
if ( (data[i]->iter=sam_itr_querys(idx, data[i]->h, conf->reg)) == 0) {
fprintf(stderr, "[E::%s] fail to parse region '%s'\n", __func__, conf->reg);
exit(1);
}
if (i == 0) tid0 = data[i]->iter->tid, beg0 = data[i]->iter->beg, end0 = data[i]->iter->end;
hts_idx_destroy(idx);
}
if (i == 0) h = h_tmp; /* save the header of first file in list */
else {
// FIXME: to check consistency
bam_hdr_destroy(h_tmp);
}
}
// allocate data storage proportionate to number of samples being studied sm->n
gplp.n = sm->n;
gplp.n_plp = calloc(sm->n, sizeof(int));
gplp.m_plp = calloc(sm->n, sizeof(int));
gplp.plp = calloc(sm->n, sizeof(bam_pileup1_t*));
fprintf(stderr, "[%s] %d samples in %d input files\n", __func__, sm->n, n);
// write the VCF header
if (conf->flag & MPLP_BCF)
{
const char *mode;
if ( conf->flag & MPLP_VCF )
mode = (conf->flag&MPLP_NO_COMP)? "wu" : "wz"; // uncompressed VCF or compressed VCF
else
mode = (conf->flag&MPLP_NO_COMP)? "wub" : "wb"; // uncompressed BCF or compressed BCF
bcf_fp = bcf_open(conf->output_fname? conf->output_fname : "-", mode);
if (bcf_fp == NULL) {
fprintf(stderr, "[%s] failed to write to %s: %s\n", __func__, conf->output_fname? conf->output_fname : "standard output", strerror(errno));
exit(1);
}
bcf_hdr = bcf_hdr_init("w");
kstring_t str = {0,0,0};
ksprintf(&str, "##samtoolsVersion=%s+htslib-%s\n",samtools_version(),hts_version());
bcf_hdr_append(bcf_hdr, str.s);
str.l = 0;
ksprintf(&str, "##samtoolsCommand=samtools mpileup");
for (i=1; i<conf->argc; i++) ksprintf(&str, " %s", conf->argv[i]);
kputc('\n', &str);
bcf_hdr_append(bcf_hdr, str.s);
if (conf->fai_fname)
{
str.l = 0;
ksprintf(&str, "##reference=file://%s\n", conf->fai_fname);
bcf_hdr_append(bcf_hdr, str.s);
}
// todo: use/write new BAM header manipulation routines, fill also UR, M5
for (i=0; i<h->n_targets; i++)
{
str.l = 0;
ksprintf(&str, "##contig=<ID=%s,length=%d>", h->target_name[i], h->target_len[i]);
bcf_hdr_append(bcf_hdr, str.s);
}
free(str.s);
bcf_hdr_append(bcf_hdr,"##ALT=<ID=X,Description=\"Represents allele(s) other than observed.\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=INDEL,Number=0,Type=Flag,Description=\"Indicates that the variant is an INDEL.\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=IDV,Number=1,Type=Integer,Description=\"Maximum number of reads supporting an indel\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=IMF,Number=1,Type=Float,Description=\"Maximum fraction of reads supporting an indel\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=DP,Number=1,Type=Integer,Description=\"Raw read depth\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=VDB,Number=1,Type=Float,Description=\"Variant Distance Bias for filtering splice-site artefacts in RNA-seq data (bigger is better)\",Version=\"3\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=RPB,Number=1,Type=Float,Description=\"Mann-Whitney U test of Read Position Bias (bigger is better)\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=MQB,Number=1,Type=Float,Description=\"Mann-Whitney U test of Mapping Quality Bias (bigger is better)\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=BQB,Number=1,Type=Float,Description=\"Mann-Whitney U test of Base Quality Bias (bigger is better)\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=MQSB,Number=1,Type=Float,Description=\"Mann-Whitney U test of Mapping Quality vs Strand Bias (bigger is better)\">");
#if CDF_MWU_TESTS
bcf_hdr_append(bcf_hdr,"##INFO=<ID=RPB2,Number=1,Type=Float,Description=\"Mann-Whitney U test of Read Position Bias [CDF] (bigger is better)\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=MQB2,Number=1,Type=Float,Description=\"Mann-Whitney U test of Mapping Quality Bias [CDF] (bigger is better)\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=BQB2,Number=1,Type=Float,Description=\"Mann-Whitney U test of Base Quality Bias [CDF] (bigger is better)\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=MQSB2,Number=1,Type=Float,Description=\"Mann-Whitney U test of Mapping Quality vs Strand Bias [CDF] (bigger is better)\">");
#endif
bcf_hdr_append(bcf_hdr,"##INFO=<ID=SGB,Number=1,Type=Float,Description=\"Segregation based metric.\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=MQ0F,Number=1,Type=Float,Description=\"Fraction of MQ0 reads (smaller is better)\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=I16,Number=16,Type=Float,Description=\"Auxiliary tag used for calling, see description of bcf_callret1_t in bam2bcf.h\">");
bcf_hdr_append(bcf_hdr,"##INFO=<ID=QS,Number=R,Type=Float,Description=\"Auxiliary tag used for calling\">");
bcf_hdr_append(bcf_hdr,"##FORMAT=<ID=PL,Number=G,Type=Integer,Description=\"List of Phred-scaled genotype likelihoods\">");
if ( conf->fmt_flag&B2B_FMT_DP )
bcf_hdr_append(bcf_hdr,"##FORMAT=<ID=DP,Number=1,Type=Integer,Description=\"Number of high-quality bases\">");
if ( conf->fmt_flag&B2B_FMT_DV )
bcf_hdr_append(bcf_hdr,"##FORMAT=<ID=DV,Number=1,Type=Integer,Description=\"Number of high-quality non-reference bases\">");
if ( conf->fmt_flag&B2B_FMT_DPR )
bcf_hdr_append(bcf_hdr,"##FORMAT=<ID=DPR,Number=R,Type=Integer,Description=\"Number of high-quality bases observed for each allele\">");
if ( conf->fmt_flag&B2B_INFO_DPR )
bcf_hdr_append(bcf_hdr,"##INFO=<ID=DPR,Number=R,Type=Integer,Description=\"Number of high-quality bases observed for each allele\">");
if ( conf->fmt_flag&B2B_FMT_DP4 )
bcf_hdr_append(bcf_hdr,"##FORMAT=<ID=DP4,Number=4,Type=Integer,Description=\"Number of high-quality ref-fwd, ref-reverse, alt-fwd and alt-reverse bases\">");
if ( conf->fmt_flag&B2B_FMT_SP )
bcf_hdr_append(bcf_hdr,"##FORMAT=<ID=SP,Number=1,Type=Integer,Description=\"Phred-scaled strand bias P-value\">");
for (i=0; i<sm->n; i++)
bcf_hdr_add_sample(bcf_hdr, sm->smpl[i]);
bcf_hdr_add_sample(bcf_hdr, NULL);
bcf_hdr_write(bcf_fp, bcf_hdr);
bca = bcf_call_init(-1., conf->min_baseQ);
bcr = calloc(sm->n, sizeof(bcf_callret1_t));
bca->rghash = rghash;
bca->openQ = conf->openQ, bca->extQ = conf->extQ, bca->tandemQ = conf->tandemQ;
bca->min_frac = conf->min_frac;
bca->min_support = conf->min_support;
bca->per_sample_flt = conf->flag & MPLP_PER_SAMPLE;
bc.bcf_hdr = bcf_hdr;
bc.n = sm->n;
bc.PL = malloc(15 * sm->n * sizeof(*bc.PL));
if (conf->fmt_flag)
{
assert( sizeof(float)==sizeof(int32_t) );
bc.DP4 = malloc(sm->n * sizeof(int32_t) * 4);
bc.fmt_arr = malloc(sm->n * sizeof(float)); // all fmt_flag fields
if ( conf->fmt_flag&(B2B_INFO_DPR|B2B_FMT_DPR) )
{
// first B2B_MAX_ALLELES fields for total numbers, the rest per-sample
bc.DPR = malloc((sm->n+1)*B2B_MAX_ALLELES*sizeof(int32_t));
for (i=0; i<sm->n; i++)
bcr[i].DPR = bc.DPR + (i+1)*B2B_MAX_ALLELES;
}
}
}
else {
pileup_fp = conf->output_fname? fopen(conf->output_fname, "w") : stdout;
if (pileup_fp == NULL) {
fprintf(stderr, "[%s] failed to write to %s: %s\n", __func__, conf->output_fname, strerror(errno));
exit(1);
}
}
if (tid0 >= 0 && conf->fai) { // region is set
ref = faidx_fetch_seq(conf->fai, h->target_name[tid0], 0, 0x7fffffff, &ref_len);
ref_tid = tid0;
for (i = 0; i < n; ++i) data[i]->ref = ref, data[i]->ref_id = tid0;
} else ref_tid = -1, ref = 0;
// begin pileup
iter = bam_mplp_init(n, mplp_func, (void**)data);
if ( conf->flag & MPLP_SMART_OVERLAPS ) bam_mplp_init_overlaps(iter);
max_depth = conf->max_depth;
if (max_depth * sm->n > 1<<20)
fprintf(stderr, "(%s) Max depth is above 1M. Potential memory hog!\n", __func__);
if (max_depth * sm->n < 8000) {
max_depth = 8000 / sm->n;
fprintf(stderr, "<%s> Set max per-file depth to %d\n", __func__, max_depth);
}
max_indel_depth = conf->max_indel_depth * sm->n;
bam_mplp_set_maxcnt(iter, max_depth);
bcf1_t *bcf_rec = bcf_init1();
int ret;
while ( (ret=bam_mplp_auto(iter, &tid, &pos, n_plp, plp)) > 0) {
if (conf->reg && (pos < beg0 || pos >= end0)) continue; // out of the region requested
if (conf->bed && tid >= 0 && !bed_overlap(conf->bed, h->target_name[tid], pos, pos+1)) continue;
if (tid != ref_tid) {
free(ref); ref = 0;
if (conf->fai) ref = faidx_fetch_seq(conf->fai, h->target_name[tid], 0, 0x7fffffff, &ref_len);
for (i = 0; i < n; ++i) data[i]->ref = ref, data[i]->ref_id = tid;
ref_tid = tid;
}
if (conf->flag & MPLP_BCF) {
int total_depth, _ref0, ref16;
for (i = total_depth = 0; i < n; ++i) total_depth += n_plp[i];
group_smpl(&gplp, sm, &buf, n, fn, n_plp, plp, conf->flag & MPLP_IGNORE_RG);
_ref0 = (ref && pos < ref_len)? ref[pos] : 'N';
ref16 = seq_nt16_table[_ref0];
bcf_callaux_clean(bca, &bc);
for (i = 0; i < gplp.n; ++i)
bcf_call_glfgen(gplp.n_plp[i], gplp.plp[i], ref16, bca, bcr + i);
bc.tid = tid; bc.pos = pos;
bcf_call_combine(gplp.n, bcr, bca, ref16, &bc);
bcf_clear1(bcf_rec);
bcf_call2bcf(&bc, bcf_rec, bcr, conf->fmt_flag, 0, 0);
bcf_write1(bcf_fp, bcf_hdr, bcf_rec);
// call indels; todo: subsampling with total_depth>max_indel_depth instead of ignoring?
if (!(conf->flag&MPLP_NO_INDEL) && total_depth < max_indel_depth && bcf_call_gap_prep(gplp.n, gplp.n_plp, gplp.plp, pos, bca, ref, rghash) >= 0)
{
bcf_callaux_clean(bca, &bc);
for (i = 0; i < gplp.n; ++i)
bcf_call_glfgen(gplp.n_plp[i], gplp.plp[i], -1, bca, bcr + i);
if (bcf_call_combine(gplp.n, bcr, bca, -1, &bc) >= 0) {
bcf_clear1(bcf_rec);
bcf_call2bcf(&bc, bcf_rec, bcr, conf->fmt_flag, bca, ref);
bcf_write1(bcf_fp, bcf_hdr, bcf_rec);
}
}
} else {
fprintf(pileup_fp, "%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, cnt;
for (j = cnt = 0; j < n_plp[i]; ++j) {
const bam_pileup1_t *p = plp[i] + j;
if (bam_get_qual(p->b)[p->qpos] >= conf->min_baseQ) ++cnt;
}
fprintf(pileup_fp, "\t%d\t", cnt);
if (n_plp[i] == 0) {
fputs("*\t*", pileup_fp);
if (conf->flag & MPLP_PRINT_MAPQ) fputs("\t*", pileup_fp);
if (conf->flag & MPLP_PRINT_POS) fputs("\t*", pileup_fp);
} else {
for (j = 0; j < n_plp[i]; ++j) {
const bam_pileup1_t *p = plp[i] + j;
if (bam_get_qual(p->b)[p->qpos] >= conf->min_baseQ)
pileup_seq(pileup_fp, plp[i] + j, pos, ref_len, ref);
}
putc('\t', pileup_fp);
for (j = 0; j < n_plp[i]; ++j) {
const bam_pileup1_t *p = plp[i] + j;
int c = bam_get_qual(p->b)[p->qpos];
if (c >= conf->min_baseQ) {
c = c + 33 < 126? c + 33 : 126;
putc(c, pileup_fp);
}
}
if (conf->flag & MPLP_PRINT_MAPQ) {
putc('\t', pileup_fp);
for (j = 0; j < n_plp[i]; ++j) {
const bam_pileup1_t *p = plp[i] + j;
int c = bam_get_qual(p->b)[p->qpos];
if ( c < conf->min_baseQ ) continue;
c = plp[i][j].b->core.qual + 33;
if (c > 126) c = 126;
putc(c, pileup_fp);
}
}
if (conf->flag & MPLP_PRINT_POS) {
putc('\t', pileup_fp);
for (j = 0; j < n_plp[i]; ++j) {
if (j > 0) putc(',', pileup_fp);
fprintf(pileup_fp, "%d", plp[i][j].qpos + 1); // FIXME: printf() is very slow...
}
}
}
}
putc('\n', pileup_fp);
}
}
// clean up
free(bc.tmp.s);
bcf_destroy1(bcf_rec);
if (bcf_fp)
{
hts_close(bcf_fp);
bcf_hdr_destroy(bcf_hdr);
bcf_call_destroy(bca);
free(bc.PL);
free(bc.DP4);
free(bc.DPR);
free(bc.fmt_arr);
free(bcr);
}
if (pileup_fp && conf->output_fname) fclose(pileup_fp);
bam_smpl_destroy(sm); free(buf.s);
for (i = 0; i < gplp.n; ++i) free(gplp.plp[i]);
free(gplp.plp); free(gplp.n_plp); free(gplp.m_plp);
bcf_call_del_rghash(rghash);
bam_mplp_destroy(iter);
bam_hdr_destroy(h);
for (i = 0; i < n; ++i) {
sam_close(data[i]->fp);
if (data[i]->iter) hts_itr_destroy(data[i]->iter);
free(data[i]);
}
free(data); free(plp); free(ref); free(n_plp);
return ret;
}
void abcWriteBcf::print(funkyPars *pars){
if(doBcf==0)
return;
kstring_t buf;
if(fp==NULL){
buf.s=NULL;buf.l=buf.m=0;
fp=aio::openFileHts(outfiles,".bcf");
hdr = bcf_hdr_init("w");
rec = bcf_init1();
print_bcf_header(fp,hdr,args,buf,header);
}
lh3struct *lh3 = (lh3struct*) pars->extras[5];
freqStruct *freq = (freqStruct *) pars->extras[6];
genoCalls *geno = (genoCalls *) pars->extras[10];
for(int s=0;s<pars->numSites;s++){
if(pars->keepSites[s]==0)
continue;
rec->rid = bcf_hdr_name2id(hdr,header->target_name[pars->refId]);
rec->pos = pars->posi[s];//<- maybe one index?
// bcf_update_id(hdr, rec, "rs6054257");
char majmin[4]={intToRef[pars->major[s]],',',intToRef[pars->minor[s]],'\0'};
bcf_update_alleles_str(hdr, rec, majmin);
rec->qual = 29;
// .. FILTER
int32_t tmpi = bcf_hdr_id2int(hdr, BCF_DT_ID, "PASS");
bcf_update_filter(hdr, rec, &tmpi, 1);
// .. INFO
tmpi = pars->keepSites[s];
bcf_update_info_int32(hdr, rec, "NS", &tmpi, 1);
if(pars->counts){
int depth = 0;
for(int i=0; i<4*pars->nInd; i++)
depth += pars->counts[s][i];
tmpi = depth;
bcf_update_info_int32(hdr, rec, "DP", &tmpi, 1);
}
if(freq){
float tmpf = freq->freq_EM[s];
bcf_update_info_float(hdr, rec, "AF", &tmpf, 1);
}
// .. FORMAT
assert(geno);
if(geno){
int32_t *tmpia = (int*)malloc(bcf_hdr_nsamples(hdr)*2*sizeof(int32_t));
for(int i=0; i<pars->nInd;i++){
if(geno->dat[s][i]==0){
tmpia[2*i+0] = bcf_gt_unphased(0);
tmpia[2*i+1] = bcf_gt_unphased(0);
}else if(geno->dat[s][i]==1){
tmpia[2*i+0] = bcf_gt_unphased(0);
tmpia[2*i+1] = bcf_gt_unphased(1);
} else{
tmpia[2*i+0] = bcf_gt_unphased(1);
tmpia[2*i+1] = bcf_gt_unphased(1);
}
}
bcf_update_genotypes(hdr, rec, tmpia, bcf_hdr_nsamples(hdr)*2);
free(tmpia);
}
if(pars->counts){
int32_t *tmpfa = (int32_t*)malloc(sizeof(int32_t)*bcf_hdr_nsamples(hdr));
suint *ary=pars->counts[s];
for(int i=0;i<bcf_hdr_nsamples(hdr);i++)
tmpfa[i] = ary[0]+ary[1]+ary[2]+ary[3];
bcf_update_format_int32(hdr, rec, "DP", tmpfa,bcf_hdr_nsamples(hdr) );
free(tmpfa);
}
assert(lh3);
if(lh3){
float *tmpfa = (float*)malloc(3*bcf_hdr_nsamples(hdr)*sizeof(float ));
int32_t *tmpi = (int32_t*)malloc(3*bcf_hdr_nsamples(hdr)*sizeof(int32_t));
double *ary = lh3->lh3[s];
for(int i=0;i<bcf_hdr_nsamples(hdr);i++)
for(int j=0;j<3;j++){
tmpfa[i*3+j] = ary[i*3+j]/M_LN10;
tmpi[i*3+j] =(int) -log10(exp(ary[i*3+j]))*10.0;
// fprintf(stderr,"pl:%d raw:%f\n",tmpi[i*3+j],ary[i*3+j]);
}
bcf_update_format_float(hdr, rec, "GL", tmpfa,3*bcf_hdr_nsamples(hdr) );
bcf_update_format_int32(hdr, rec, "PL", tmpi,3*bcf_hdr_nsamples(hdr) );
free(tmpfa);
free(tmpi);
}
if ( bcf_write1(fp, hdr, rec)!=0 ){
fprintf(stderr,"Failed to write to \n");
exit(0);
}
// fprintf(stderr,"------\n");
bcf_clear1(rec);
}
}
static void init_data(args_t *args)
{
args->aux.srs = bcf_sr_init();
// Open files for input and output, initialize structures
if ( args->targets )
{
if ( bcf_sr_set_targets(args->aux.srs, args->targets, args->targets_is_file, args->aux.flag&CALL_CONSTR_ALLELES ? 3 : 0)<0 )
error("Failed to read the targets: %s\n", args->targets);
if ( args->aux.flag&CALL_CONSTR_ALLELES && args->flag&CF_INS_MISSED )
{
args->aux.srs->targets->missed_reg_handler = print_missed_line;
args->aux.srs->targets->missed_reg_data = args;
}
}
if ( args->regions )
{
if ( bcf_sr_set_regions(args->aux.srs, args->regions, args->regions_is_file)<0 )
error("Failed to read the targets: %s\n", args->regions);
}
int i;
if ( !bcf_sr_add_reader(args->aux.srs, args->bcf_fname) ) error("Failed to open: %s\n", args->bcf_fname);
if ( args->nsamples && args->nsamples != bcf_hdr_nsamples(args->aux.srs->readers[0].header) )
{
args->samples_map = (int *) malloc(sizeof(int)*args->nsamples);
args->aux.hdr = bcf_hdr_subset(args->aux.srs->readers[0].header, args->nsamples, args->samples, args->samples_map);
for (i=0; i<args->nsamples; i++)
if ( args->samples_map[i]<0 ) error("No such sample: %s\n", args->samples[i]);
if ( !bcf_hdr_nsamples(args->aux.hdr) ) error("No matching sample found\n");
}
else
{
args->aux.hdr = bcf_hdr_dup(args->aux.srs->readers[0].header);
for (i=0; i<args->nsamples; i++)
if ( bcf_hdr_id2int(args->aux.hdr,BCF_DT_SAMPLE,args->samples[i])<0 )
error("No such sample: %s\n", args->samples[i]);
}
// Reorder ploidy and family indexes to match mpileup's output and exclude samples which are not available
if ( args->aux.ploidy )
{
for (i=0; i<args->aux.nfams; i++)
{
int j;
for (j=0; j<3; j++)
{
int k = bcf_hdr_id2int(args->aux.hdr, BCF_DT_SAMPLE, args->samples[ args->aux.fams[i].sample[j] ]);
if ( k<0 ) error("No such sample: %s\n", args->samples[ args->aux.fams[i].sample[j] ]);
args->aux.fams[i].sample[j] = k;
}
}
uint8_t *ploidy = (uint8_t*) calloc(bcf_hdr_nsamples(args->aux.hdr), 1);
for (i=0; i<args->nsamples; i++) // i index in -s sample list
{
int j = bcf_hdr_id2int(args->aux.hdr, BCF_DT_SAMPLE, args->samples[i]); // j index in the output VCF / subset VCF
if ( j<0 )
{
fprintf(stderr,"Warning: no such sample: \"%s\"\n", args->samples[i]);
continue;
}
ploidy[j] = args->aux.ploidy[i];
}
args->nsamples = bcf_hdr_nsamples(args->aux.hdr);
for (i=0; i<args->nsamples; i++)
assert( ploidy[i]==0 || ploidy[i]==1 || ploidy[i]==2 );
free(args->aux.ploidy);
args->aux.ploidy = ploidy;
}
args->out_fh = hts_open(args->output_fname, hts_bcf_wmode(args->output_type));
if ( args->out_fh == NULL ) error("Can't write to \"%s\": %s\n", args->output_fname, strerror(errno));
if ( args->flag & CF_QCALL )
return;
if ( args->flag & CF_MCALL )
mcall_init(&args->aux);
if ( args->flag & CF_CCALL )
ccall_init(&args->aux);
if ( args->flag&CF_GVCF )
{
bcf_hdr_append(args->aux.hdr,"##INFO=<ID=END,Number=1,Type=Integer,Description=\"End position of the variant described in this record\">");
args->gvcf.rid = -1;
args->gvcf.line = bcf_init1();
args->gvcf.gt = (int32_t*) malloc(2*sizeof(int32_t)*bcf_hdr_nsamples(args->aux.hdr));
for (i=0; i<bcf_hdr_nsamples(args->aux.hdr); i++)
{
args->gvcf.gt[2*i+0] = bcf_gt_unphased(0);
args->gvcf.gt[2*i+1] = bcf_gt_unphased(0);
}
}
bcf_hdr_remove(args->aux.hdr, BCF_HL_INFO, "QS");
bcf_hdr_remove(args->aux.hdr, BCF_HL_INFO, "I16");
bcf_hdr_append_version(args->aux.hdr, args->argc, args->argv, "bcftools_call");
bcf_hdr_write(args->out_fh, args->aux.hdr);
if ( args->flag&CF_INS_MISSED ) init_missed_line(args);
}
static void init_data(args_t *args)
{
args->aux.srs = bcf_sr_init();
// Open files for input and output, initialize structures
if ( args->targets )
{
if ( bcf_sr_set_targets(args->aux.srs, args->targets, args->targets_is_file, args->aux.flag&CALL_CONSTR_ALLELES ? 3 : 0)<0 )
error("Failed to read the targets: %s\n", args->targets);
if ( args->aux.flag&CALL_CONSTR_ALLELES && args->flag&CF_INS_MISSED )
{
args->aux.srs->targets->missed_reg_handler = print_missed_line;
args->aux.srs->targets->missed_reg_data = args;
}
}
if ( args->regions )
{
if ( bcf_sr_set_regions(args->aux.srs, args->regions, args->regions_is_file)<0 )
error("Failed to read the targets: %s\n", args->regions);
}
if ( !bcf_sr_add_reader(args->aux.srs, args->bcf_fname) ) error("Failed to open %s: %s\n", args->bcf_fname,bcf_sr_strerror(args->aux.srs->errnum));
args->aux.hdr = bcf_sr_get_header(args->aux.srs,0);
int i;
if ( args->samples_fname )
{
set_samples(args, args->samples_fname, args->samples_is_file);
if ( args->aux.flag&CALL_CONSTR_TRIO )
{
if ( 3*args->aux.nfams!=args->nsamples ) error("Expected only trios in %s, sorry!\n", args->samples_fname);
fprintf(stderr,"Detected %d samples in %d trio families\n", args->nsamples,args->aux.nfams);
}
args->nsex = ploidy_nsex(args->ploidy);
args->sex2ploidy = (int*) calloc(args->nsex,sizeof(int));
args->sex2ploidy_prev = (int*) calloc(args->nsex,sizeof(int));
args->aux.ploidy = (uint8_t*) malloc(args->nsamples);
for (i=0; i<args->nsamples; i++) args->aux.ploidy[i] = 2;
for (i=0; i<args->nsex; i++) args->sex2ploidy_prev[i] = 2;
}
if ( args->samples_map )
{
args->aux.hdr = bcf_hdr_subset(bcf_sr_get_header(args->aux.srs,0), args->nsamples, args->samples, args->samples_map);
if ( !args->aux.hdr ) error("Error occurred while subsetting samples\n");
for (i=0; i<args->nsamples; i++)
if ( args->samples_map[i]<0 ) error("No such sample: %s\n", args->samples[i]);
if ( !bcf_hdr_nsamples(args->aux.hdr) ) error("No matching sample found\n");
}
else
{
args->aux.hdr = bcf_hdr_dup(bcf_sr_get_header(args->aux.srs,0));
for (i=0; i<args->nsamples; i++)
if ( bcf_hdr_id2int(args->aux.hdr,BCF_DT_SAMPLE,args->samples[i])<0 )
error("No such sample: %s\n", args->samples[i]);
}
args->out_fh = hts_open(args->output_fname, hts_bcf_wmode(args->output_type));
if ( args->out_fh == NULL ) error("Can't write to \"%s\": %s\n", args->output_fname, strerror(errno));
if ( args->flag & CF_QCALL )
return;
if ( args->flag & CF_MCALL )
mcall_init(&args->aux);
if ( args->flag & CF_CCALL )
ccall_init(&args->aux);
if ( args->flag&CF_GVCF )
{
bcf_hdr_append(args->aux.hdr,"##INFO=<ID=END,Number=1,Type=Integer,Description=\"End position of the variant described in this record\">");
args->gvcf.rid = -1;
args->gvcf.line = bcf_init1();
args->gvcf.gt = (int32_t*) malloc(2*sizeof(int32_t)*bcf_hdr_nsamples(args->aux.hdr));
for (i=0; i<bcf_hdr_nsamples(args->aux.hdr); i++)
{
args->gvcf.gt[2*i+0] = bcf_gt_unphased(0);
args->gvcf.gt[2*i+1] = bcf_gt_unphased(0);
}
}
bcf_hdr_remove(args->aux.hdr, BCF_HL_INFO, "QS");
bcf_hdr_remove(args->aux.hdr, BCF_HL_INFO, "I16");
bcf_hdr_append_version(args->aux.hdr, args->argc, args->argv, "bcftools_call");
bcf_hdr_write(args->out_fh, args->aux.hdr);
if ( args->flag&CF_INS_MISSED ) init_missed_line(args);
}