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VarScan2

version=v2.3.9.a.1

  1. Modify function comparePositions() in Somatic.java

Use normalMinVarFreqDecideGenotype instead of MinVarFreq to decide Normal's genotype when Tumor and Normal are both HET(CASE 2E).

normalMinVarFreqDecideGenotype = 0.03, when MinVarFreq < 0.03, else = MinVarFreq.

It make sense when MinVarFreq is pretty low(e.g 0.005), avoid taking normal HET falsely.

This may call MORE somatic snp/indel!

The difference between my version and origin is showed below: test_data/test.snp.vcf is my version's result, the other is origin version's.

diff --unified=0 test_data/test.snp.vcf /data_sas/ypu/git_repository/Med1CDx/test_data/ZY/call/chrom/sample.chr7\:10000-159128663.snp.vcf

--- test_data/test.snp.vcf	2019-03-08 11:41:03.059672490 +0800
+++ /data_sas/ypu/git_repository/Med1CDx/test_data/ZY/call/chrom/sample.chr7:10000-159128663.snp.vcf  2019-03-05 16:02:08.653294173 +0800
@@ -1871 +1871 @@
-chr7	140453136	.	A	T	.	PASS	DP=2279;SOMATIC;SS=2;SSC=255;GPV=1E0;SPV=8.1581E-59	GT:GQ:DP:RD:AD:FREQ:DP4	0/1:.:378:372:6:1.59%:258,114,5,1	0/1:.:1901:1189:712:37.45%:1001,188,636,76
+chr7	140453136	.	A	T	.	PASS	DP=2279;SS=1;SSC=255;GPV=1E0;SPV=8.1581E-59	GT:GQ:DP:RD:AD:FREQ:DP4	0/1:.:378:372:6:1.59%:258,114,5,1	0/1:.:1901:1189:712:37.45%:1001,188,636,76

@@ -2672 +2672 @@
-chr7	151933014	.	C	T	.	PASS	DP=1922;SS=1;SSC=16;GPV=1.7461E-25;SPV=2.5102E-2	GT:GQ:DP:RD:AD:FREQ:DP4	0/1:.:383:372:9:2.36%:209,163,1,8	0/1:.:1539:1466:72:4.68%:1242,224,4,68
+chr7	151933014	.	C	T	.	PASS	DP=1922;SS=1;SSC=16;GPV=1E0;SPV=2.5102E-2	GT:GQ:DP:RD:AD:FREQ:DP4	0/1:.:383:372:9:2.36%:209,163,1,8	0/1:.:1539:1466:72:4.68%:1242,224,4,68

@@ -2681 +2681 @@
-chr7	151935689	.	T	C	.	PASS	DP=1586;SS=1;SSC=16;GPV=2.2796E-8;SPV=2.4017E-2	GT:GQ:DP:RD:AD:FREQ:DP4	0/1:.:458:201:4:1.95%:196,5,0,4	0/1:.:1128:346:21:5.72%:342,4,0,21
+chr7	151935689	.	T	C	.	PASS	DP=1586;SS=1;SSC=16;GPV=1E0;SPV=2.4017E-2	GT:GQ:DP:RD:AD:FREQ:DP4	0/1:.:458:201:4:1.95%:196,5,0,4	0/1:.:1128:346:21:5.72%:342,4,0,21
@@ -2766 +2766 @@
-chr7	151970959	.	G	A	.	PASS	DP=3510;SS=1;SSC=62;GPV=2.4658E-14;SPV=5.4694E-7	GT:GQ:DP:RD:AD:FREQ:DP4	0/1:.:1216:1181:32:2.64%:420,761,8,24	0/1:.:2294:2281:13:0.57%:1799,482,4,9
+chr7	151970959	.	G	A	.	PASS	DP=3510;SS=1;SSC=62;GPV=1E0;SPV=5.4694E-7	GT:GQ:DP:RD:AD:FREQ:DP4	0/1:.:1216:1181:32:2.64%:420,761,8,24	0/1:.:2294:2281:13:0.57%:1799,482,4,9
  1. Add a variant named freqQuot in comparePositions() in Somatic.java freqQuot= tumorFreq / normalFreq. In CASE 2E(Variant alleles match but difference significant), if freqQuot >= 10.0, we can treat this mutation as Somatic. For example tumorFreq=7.0% and normal=0.3%.

diff -u after.vcf before.vcf

-chr2    29416481        .       T       C       .       PASS    DP=2030;SOMATIC;SS=2;SSC=198;GPV=1E0;SPV=1.3145E-20     GT:GQ:DP:RD:AD:FREQ:DP4 0/1:.:1078:1073:4:0.37%:859,214,3,1     0/1:.:952:877:74:7.78%:744,133,64,10
+chr2    29416481        .       T       C       .       PASS    DP=2030;SS=1;SSC=198;GPV=1.5554E-24;SPV=1.3145E-20      GT:GQ:DP:RD:AD:FREQ:DP4 0/1:.:1078:1073:4:0.37%:859,214,3,1     0/1:.:952:877:74:7.78%:744,133,64,10

NOTE

This version NOT fix the bug: When normalReads2 = 1 < minReads2 = 2, this may make diffPvalue > 0.05. Set normalReads2 = 0, if normalAllele2.equals(refBase), which is fixed in other branch(v2.3.9.b).

  1. Fix a bug: When normalReads2 = 1 < minReads2 = 2, this may make diffPvalue > 0.05. Set normalReads2 = 0, if normalAllele2.equals(refBase).

my version:

chr7	55242464	.	AGGAATTAAGAGAAGC	A	.	PASS	DP=1819; SOMATIC;SS=2;SSC=6;GPV=1E0;SPV=2.3063E-1	GT:GQ:DP:RD:AD:FREQ:DP4	0/0:.:305:304:0:0%:172,132,1,0	0/1:.:1514:1506:8:0.53%:692,814,6,2

origin version:

chr7    55242464	.	AGGAATTAAGAGAAGC	A	.	PASS	DP=1819;SS=0;SSC=0;GPV=1E0;SPV=5.3897E-1	GT:GQ:DP:RD:AD:FREQ:DP4	0/0:.:305:304:1:0.33%:172,132,1,0	0/1:.:1514:1506:8:0.53%:692,814,6,2

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Fix some bugs for low min-variant-freq & some MS site reported as Somatic-indel issue.

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