Why is this insertion detected by NGS a false positive ?
0
2
Entering edit mode
8.1 years ago
docfak ▴ 20

Hello,

We performed whole exome sequencing on a patient with an AmpliSeq Exome kit and the IonTorrent/Life Proton sequencer. Alignment to human genome reference and variant calling are done by IonTorrent plugins. After bioinformatics analysis, we retained one insertion which was infirmed by Sanger sequencing.

I can not determine why this variant is a false positive. Could it be a alignment problem caused by the homopolymers situated some bp after the variant ?

Here is the data :

CHROM X
POS 153667132
REF C
ALT CG
QUAL 98.7837
FILTER PASS
AF 0.84375
AO 23
CG 2.000
CP 0.252
DP 33
DSP 22
FAO 27
FD unknown
FDP 32
FG frameshift
FR .
FRO 5
FSAF 21
FSAR 6
FSRF 5
FSRR 0
FWDB 0.0947456
FXX 0.0302939
GL GDI1
HRUN 0
LEN 1
MILD 25.3658
OALT G
PB 0.5
PBP 1
QD 12.348
RBI 0.232263
REFB -0.107327
REVB 0.212059
RO 8
SAF 17
SAR 6
SRF 8
SRR 0
SSEN 0
SSEP 0
SSSB -0.175701
STB 0.553155
STBP 0.288
TYPE ins
VARB 0.0524829
DT 0
HS 0

And some screens from IGV :

IGV Screenshot 1

IGV Screenshot 2

IGV Screenshot 3

Many thanks in advance for your help.

next-gen sequencing • 3.6k views
ADD COMMENT
1
Entering edit mode

From the IGV views, it appears that there's so much activity around there which usually means a poor mapping quality or base qualities along the read. I did check to see if the region was in some kind of a duplication/repeat rich region but that doesn't appear to be the case from the UCSC browser.

ADD REPLY
0
Entering edit mode

Thank you for your quick answer !

ADD REPLY
0
Entering edit mode

IonTorrent is just bad at tiny indels. I bet it is a sequencing problem rather than an informatics problem.

ADD REPLY

Login before adding your answer.

Traffic: 2455 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6