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removed contig weight & code cleanup 

Contig weight was calculated based on a false assumption. It is actually not useful.
This commit is contained in:
Heng Li 2014-11-07 12:45:01 -05:00
parent 4e97cdfde8
commit b2b42cea7e
1 changed files with 35 additions and 157 deletions
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192
bwa-postalt.js
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@ -52,31 +52,6 @@ var getopt = function(args, ostr) {
return optopt; return optopt;
} }
// print an object in a format similar to JSON. For debugging only.
function obj2str(o)
{
if (typeof(o) != 'object') {
return o.toString();
} else if (o == null) {
return "null";
} else if (Array.isArray(o)) {
var s = "[";
for (var i = 0; i < o.length; ++i) {
if (i) s += ',';
s += obj2str(o[i]);
}
return s + "]";
} else {
var i = 0, s = "{";
for (var key in o) {
if (i++) s += ',';
s += key + ":";
s += obj2str(o[key]);
}
return s + "}";
}
}
// reverse a string // reverse a string
Bytes.prototype.reverse = function() Bytes.prototype.reverse = function()
{ {
@ -219,32 +194,19 @@ function print_buffer(buf2, fp_hla, hla)
function bwa_postalt(args) function bwa_postalt(args)
{ {
var c, opt = { a:1, b:4, o:6, e:1, verbose:3, show_pri:false, update_mapq:true, min_mapq:10, min_sc:90, max_nm_sc:10, show_ev:false, min_pa_ratio:1 }; var c, opt = { a:1, b:4, o:6, e:1, min_mapq:10, min_sc:90, max_nm_sc:10, min_pa_ratio:1 };
while ((c = getopt(args, 'Pqev:p:r:')) != null) { while ((c = getopt(args, 'p:r:')) != null) {
if (c == 'v') opt.verbose = parseInt(getopt.arg); if (c == 'p') opt.pre = getopt.arg;
else if (c == 'p') opt.pre = getopt.arg;
else if (c == 'P') opt.show_pri = true;
else if (c == 'q') opt.recover_maq = false;
else if (c == 'e') opt.show_ev = true;
else if (c == 'r') opt.min_pa_ratio = parseFloat(getopt.arg); else if (c == 'r') opt.min_pa_ratio = parseFloat(getopt.arg);
} }
if (opt.min_pa_ratio > 1.) opt.min_pa_ratio = 1.; if (opt.min_pa_ratio > 1.) opt.min_pa_ratio = 1.;
if (opt.show_ev && opt.pre == null) {
warn("ERROR: option '-p' must be specified if '-e' is applied.");
exit(1);
}
if (args.length == getopt.ind) { if (args.length == getopt.ind) {
print(""); print("");
print("Usage: k8 bwa-postalt.js [options] <alt.sam> [aln.sam]\n"); print("Usage: k8 bwa-postalt.js [options] <alt.sam> [aln.sam]\n");
print("Options: -p STR prefix of file(s) for additional information [null]"); print("Options: -p STR prefix of files matching HLA genes [null]");
print(" PREFIX.ctw - weight of each ALT contig");
print(" PREFIX.evi - reads supporting ALT contigs (effective with -e)");
print(" -e show reads supporting ALT contigs into file PREFIX.evi");
print(" -r FLOAT reduce mapQ to 0 if not overlapping lifted best and pa<FLOAT ["+opt.min_pa_ratio+"]"); print(" -r FLOAT reduce mapQ to 0 if not overlapping lifted best and pa<FLOAT ["+opt.min_pa_ratio+"]");
print(" -q don't modify mapQ for non-ALTs hit overlapping lifted ALT");
print(""); print("");
print("Note: This script extracts the XA tag, lifts the mapping positions of ALT hits to"); print("Note: This script extracts the XA tag, lifts the mapping positions of ALT hits to");
print(" the primary assembly, groups them and then estimates mapQ across groups. If"); print(" the primary assembly, groups them and then estimates mapQ across groups. If");
@ -256,24 +218,21 @@ function bwa_postalt(args)
exit(1); exit(1);
} }
var fp_evi = opt.show_ev && opt.pre? new File(opt.pre + '.evi', "w") : null;
var aux = new Bytes(); // used for reverse and reverse complement var aux = new Bytes(); // used for reverse and reverse complement
var buf = new Bytes(); var buf = new Bytes(); // line reading buffer
// read ALT-to-REF alignment // read ALT-to-REF alignment
var intv_alt = {}, intv_pri = {}, idx_un = {}, hla_ctg = {}; var intv_alt = {}, intv_pri = {}, hla_ctg = {}, is_alt = {};
var file = new File(args[getopt.ind]); var file = new File(args[getopt.ind]);
while (file.readline(buf) >= 0) { while (file.readline(buf) >= 0) {
var line = buf.toString(); var line = buf.toString();
if (line.charAt(0) == '@') continue; if (line.charAt(0) == '@') continue;
var t = line.split("\t"); var t = line.split("\t");
if (t.length < 11) continue; // incomplete lines if (t.length < 11) continue; // incomplete lines
is_alt[t[0]] = true;
var pos = parseInt(t[3]) - 1; var pos = parseInt(t[3]) - 1;
var flag = parseInt(t[1]); var flag = parseInt(t[1]);
if ((flag&4) || t[2] == '*') { if ((flag&4) || t[2] == '*') continue;
idx_un[t[0]] = true;
continue;
}
var m, cigar = [], l_qaln = 0, l_tlen = 0, l_qclip = 0; var m, cigar = [], l_qaln = 0, l_tlen = 0, l_qclip = 0;
if ((m = /^(HLA-[^\s\*]+)\*\d+/.exec(t[0])) != null) { // read HLA contigs if ((m = /^(HLA-[^\s\*]+)\*\d+/.exec(t[0])) != null) { // read HLA contigs
if (hla_ctg[m[1]] == null) hla_ctg[m[1]] = 0; if (hla_ctg[m[1]] == null) hla_ctg[m[1]] = 0;
@ -296,10 +255,8 @@ function bwa_postalt(args)
} }
file.close(); file.close();
var idx_alt = {}, idx_pri = {}; var idx_alt = {}, idx_pri = {};
for (var ctg in intv_alt) for (var ctg in intv_alt) idx_alt[ctg] = intv_ovlp(intv_alt[ctg]);
idx_alt[ctg] = intv_ovlp(intv_alt[ctg]); for (var ctg in intv_pri) idx_pri[ctg] = intv_ovlp(intv_pri[ctg]);
for (var ctg in intv_pri)
idx_pri[ctg] = intv_ovlp(intv_pri[ctg]);
// initialize the list of HLA contigs // initialize the list of HLA contigs
var fp_hla = null; var fp_hla = null;
@ -309,18 +266,12 @@ function bwa_postalt(args)
fp_hla[h] = new File(opt.pre + '.' + h + '.fq', "w"); fp_hla[h] = new File(opt.pre + '.' + h + '.fq', "w");
} }
// initialize the list of ALT contigs
var weight_alt = [];
for (var ctg in idx_alt)
weight_alt[ctg] = [0, 0, 0, 0, 0, 0, intv_alt[ctg][0][3], intv_alt[ctg][0][5], intv_alt[ctg][0][7]];
for (var ctg in idx_un)
weight_alt[ctg] = [0, 0, 0, 0, 0, 0, '~', 0, 0];
// process SAM // process SAM
var buf2 = [], hla = {}; var buf2 = [], hla = {};
file = args.length - getopt.ind >= 2? new File(args[getopt.ind+1]) : new File(); file = args.length - getopt.ind >= 2? new File(args[getopt.ind+1]) : new File();
while (file.readline(buf) >= 0) { while (file.readline(buf) >= 0) {
var m, line = buf.toString(); var m, line = buf.toString();
if (line.charAt(0) == '@') { // print and then skip the header line if (line.charAt(0) == '@') { // print and then skip the header line
print(line); print(line);
continue; continue;
@ -362,7 +313,7 @@ function bwa_postalt(args)
// check if there are ALT hits // check if there are ALT hits
var has_alt = false; var has_alt = false;
for (var i = 0; i < hits.length; ++i) for (var i = 0; i < hits.length; ++i)
if (weight_alt[hits[i].ctg] != null) { if (is_alt[hits[i].ctg] != null) {
has_alt = true; has_alt = true;
break; break;
} }
@ -429,7 +380,7 @@ function bwa_postalt(args)
if (hits[i].g == reported_g) if (hits[i].g == reported_g)
++n_group0; ++n_group0;
} else { } else {
if (weight_alt[hits[0].ctg] == null) { // no need to go through the following if the single hit is non-ALT if (is_alt[hits[0].ctg] == null) { // no need to go through the following if the single hit is non-ALT
buf2.push(t); buf2.push(t);
continue; continue;
} }
@ -455,7 +406,8 @@ function bwa_postalt(args)
else mapQ = mapQ > ori_mapQ? mapQ : ori_mapQ; else mapQ = mapQ > ori_mapQ? mapQ : ori_mapQ;
} else mapQ = t[4]; } else mapQ = t[4];
var pri_ofunc = idx_pri[hits[reported_i].pctg], ovlp_alt = []; // find out whether the read is overlapping HLA genes
var pri_ofunc = idx_pri[hits[reported_i].pctg];
if (pri_ofunc != null) { if (pri_ofunc != null) {
var rpt_start = 1<<30, rpt_end = 0; var rpt_start = 1<<30, rpt_end = 0;
for (var i = 0; i < hits.length; ++i) { for (var i = 0; i < hits.length; ++i) {
@ -465,70 +417,29 @@ function bwa_postalt(args)
rpt_end = rpt_end > h.pend ? rpt_end : h.pend; rpt_end = rpt_end > h.pend ? rpt_end : h.pend;
} }
} }
ovlp_alt = pri_ofunc(rpt_start, rpt_end); var ovlp_alt = pri_ofunc(rpt_start, rpt_end);
for (var i = 0; i < ovlp_alt.length; ++i) for (var i = 0; i < ovlp_alt.length; ++i)
if ((m = /^(HLA-[^\s\*]+)\*\d+/.exec(ovlp_alt[i][2])) != null) if ((m = /^(HLA-[^\s\*]+)\*\d+/.exec(ovlp_alt[i][2])) != null)
hla[m[1]] = true; hla[m[1]] = true;
} }
// ALT genotyping // adjust the mapQ of the primary hits
if (mapQ >= opt.min_mapq && hits[reported_i].score >= opt.min_sc) { if (n_rpt_lifted <= 1) {
// collect all overlapping ALT contigs
var alts = {};
for (var i = 0; i < hits.length; ++i) {
var h = hits[i];
if (h.g == reported_g && weight_alt[h.ctg] != null)
alts[h.ctg] = [h.score, h.NM];
}
if (ovlp_alt.length > 0) { // add other unreported hits
for (var i = 0; i < ovlp_alt.length; ++i)
if (ovlp_alt[i][0] <= rpt_start && rpt_end <= ovlp_alt[i][1] && alts[ovlp_alt[i][2]] == null)
alts[ovlp_alt[i][2]] = [0, 0];
}
// add weight to each ALT contig
var alt_arr = [], max_sc = -1, max_i = -1, sum = 0, min_sc = 1<<30, max_nm = -1;
for (var ctg in alts)
alt_arr.push([ctg, alts[ctg][0], 0, alts[ctg][1]]);
for (var i = 0; i < alt_arr.length; ++i) {
if (max_sc < alt_arr[i][1])
max_sc = alt_arr[i][1], max_i = i;
min_sc = min_sc < alt_arr[i][1]? min_sc : alt_arr[i][1];
var nm = alt_arr[i][1] > 0? alt_arr[i][3] : opt.max_nm_sc;
max_nm = max_nm > nm? max_nm : nm;
}
if (max_nm > opt.max_nm_sc) max_nm = opt.max_nm_sc;
if (max_sc > 0 && (alt_arr.length == 1 || min_sc < max_sc)) {
for (var i = 0; i < alt_arr.length; ++i)
sum += (alt_arr[i][2] = Math.pow(10, .6 * (alt_arr[i][1] - max_sc)));
for (var i = 0; i < alt_arr.length; ++i) alt_arr[i][2] /= sum;
for (var i = 0; i < alt_arr.length; ++i) {
var e = [alt_arr[i][0], 1, alt_arr[max_i][2], alt_arr[i][2], max_nm, max_nm * alt_arr[max_i][2], max_nm * alt_arr[i][2]];
var w = weight_alt[e[0]];
for (var j = 0; j < 6; ++j) w[j] += e[j+1];
if (fp_evi) {
e[2] = e[2].toFixed(3); e[3] = e[3].toFixed(3);
e[5] = e[5].toFixed(3); e[6] = e[6].toFixed(3);
fp_evi.write(t[0] + '/' + (t[1]>>6&3) + '\t' + e.join("\t") + '\n');
}
}
}
}
// check if the reported hit overlaps a hit to the primary assembly; if so, don't reduce mapping quality
if (opt.update_mapq && mapQ > 0 && n_rpt_lifted <= 1) {
var l = n_rpt_lifted == 1? rpt_lifted : null; var l = n_rpt_lifted == 1? rpt_lifted : null;
for (var i = 0; i < buf2.length; ++i) { for (var i = 0; i < buf2.length; ++i) {
var s = buf2[i], is_ovlp = true; var s = buf2[i], is_ovlp = true;
if (l != null) { if (l != null) {
if (l[0] != s[2]) is_ovlp = false; // different chr if (l[0] != s[2]) is_ovlp = false; // different chr
if (((s[1]&16) != 0) != l[1]) is_ovlp = false; // different strand else if (((s[1]&16) != 0) != l[1]) is_ovlp = false; // different strand
var start = s[3] - 1, end = start; else {
while ((m = re_cigar.exec(t[5])) != null) var start = s[3] - 1, end = start;
if (m[2] == 'M' || m[2] == 'D' || m[2] == 'N') while ((m = re_cigar.exec(t[5])) != null)
end += parseInt(m[1]); if (m[2] == 'M' || m[2] == 'D' || m[2] == 'N')
if (!(start < l[3] && l[2] < end)) is_ovlp = false; // no overlap end += parseInt(m[1]);
if (!(start < l[3] && l[2] < end)) is_ovlp = false; // no overlap
}
} else is_ovlp = false; } else is_ovlp = false;
// get the "pa" tag if present
var om = -1, pa = 10.; var om = -1, pa = 10.;
for (var j = 11; j < s.length; ++j) for (var j = 11; j < s.length; ++j)
if ((m = /^om:i:(\d+)/.exec(s[j])) != null) if ((m = /^om:i:(\d+)/.exec(s[j])) != null)
@ -555,20 +466,6 @@ function bwa_postalt(args)
} }
} }
// generate reversed quality and reverse-complemented sequence if necessary
var rs = null, rq = null; // reversed quality and reverse complement sequence
var need_rev = false;
for (var i = 0; i < hits.length; ++i) {
if (hits[i].g != reported_g || i == reported_i) continue;
if (hits[i].rev != hits[reported_i].rev)
need_rev = true;
}
if (need_rev) { // reverse and reverse complement
aux.length = 0;
aux.set(t[9], 0); aux.revcomp(); rs = aux.toString();
aux.set(t[10],0); aux.reverse(); rq = aux.toString();
}
// stage the reported hit // stage the reported hit
t[4] = mapQ; t[4] = mapQ;
if (n_group0 > 1) t.push("om:i:"+ori_mapQ); if (n_group0 > 1) t.push("om:i:"+ori_mapQ);
@ -576,18 +473,22 @@ function bwa_postalt(args)
buf2.push(t); buf2.push(t);
// stage the hits generated from the XA tag // stage the hits generated from the XA tag
var cnt = 0; var cnt = 0, rs = null, rq = null; // rq: reverse quality; rs: reverse complement sequence
var rg = (m = /\t(RG:Z:\S+)/.exec(line)) != null? m[1] : null; var rg = (m = /\t(RG:Z:\S+)/.exec(line)) != null? m[1] : null;
for (var i = 0; i < hits.length; ++i) { for (var i = 0; i < hits.length; ++i) {
if (opt.verbose >= 5) print(obj2str(hits[i]));
if (hits[i].g != reported_g || i == reported_i) continue; if (hits[i].g != reported_g || i == reported_i) continue;
if (!opt.show_pri && idx_alt[hits[i].ctg] == null) continue; if (idx_alt[hits[i].ctg] == null) continue;
var s = [t[0], 0, hits[i].ctg, hits[i].start+1, mapQ, hits[i].cigar, t[6], t[7], t[8]]; var s = [t[0], 0, hits[i].ctg, hits[i].start+1, mapQ, hits[i].cigar, t[6], t[7], t[8]];
// print sequence/quality and set the rev flag // print sequence/quality and set the rev flag
if (hits[i].rev == hits[reported_i].rev) { if (hits[i].rev == hits[reported_i].rev) {
s.push(t[9], t[10]); s.push(t[9], t[10]);
s[1] = flag | 0x800; s[1] = flag | 0x800;
} else { } else { // we need to write the reverse sequence
if (rs == null || rq == null) {
aux.length = 0;
aux.set(t[9], 0); aux.revcomp(); rs = aux.toString();
aux.set(t[10],0); aux.reverse(); rq = aux.toString();
}
s.push(rs, rq); s.push(rs, rq);
s[1] = (flag ^ 0x10) | 0x800; s[1] = (flag ^ 0x10) | 0x800;
} }
@ -599,35 +500,12 @@ function bwa_postalt(args)
} }
print_buffer(buf2, fp_hla, hla); print_buffer(buf2, fp_hla, hla);
file.close(); file.close();
if (fp_evi != null) fp_evi.close();
if (fp_hla != null) if (fp_hla != null)
for (var h in fp_hla) for (var h in fp_hla)
fp_hla[h].close(); fp_hla[h].close();
buf.destroy(); buf.destroy();
aux.destroy(); aux.destroy();
// print weight of each contig
if (opt.pre != null) {
var fpout = new File(opt.pre + '.ctw', "w");
var weight_arr = [];
var weight_hla = [];
for (var ctg in weight_alt) {
var w = weight_alt[ctg];
weight_arr.push([ctg, w[6], w[7], w[8],
w[0], w[1].toFixed(3), w[2].toFixed(3), w[1] > 0? (w[2]/w[1]).toFixed(3) : '0.000',
w[3], w[4].toFixed(3), w[5].toFixed(3), w[4] > 0? (w[5]/w[4]).toFixed(3) : '0.000']);
weight_hla.push([ctg, w[4] > 0? w[5]/w[4] : 0]);
}
weight_arr.sort(function(a,b) {
return a[1] < b[1]? -1 : a[1] > b[1]? 1 : a[2] != b[2]? a[2] - b[2] : a[0] < b[0]? -1 : a[0] > b[0]? 1 : 0;
});
for (var i = 0; i < weight_arr.length; ++i) {
if (weight_arr[i][1] == '~') weight_arr[i][1] = '*';
fpout.write(weight_arr[i].join("\t") + '\n');
}
fpout.close();
}
} }
bwa_postalt(arguments); bwa_postalt(arguments);