Detection of BRAF V600E mutation by pyrosequencing.

Abstract:

INTRODUCTION:Detection of the V600E hotspot mutation in BRAF oncogene is extremely useful for the screening of hereditary non-polyposis colorectal cancer (Lynch's syndrome) and for the prediction of sensitivity to MEK inhibitors. Here we describe a method for detecting this mutation based upon pyrosequencing technology. METHODS:The efficiency of pyrosequencing for detecting BRAF V600E mutations was compared with the conventional dideoxy sequencing method in 12 tumour cell lines and in 108 colorectal tumours. RESULTS:The results from pyrosequencing were 100% concordant with those from dideoxy sequencing. This method was capable of detecting BRAF V600E mutations at a much lower ratio of mutant to wild-type alleles (1:50) than dideoxy sequencing (1:5) while being considerably faster and less expensive. CONCLUSIONS:Pyrosequencing offers a specific, sensitive, rapid and cost-effective alternative to dideoxy sequencing for the detection of BRAF V600E mutations in clinical tumour specimens.

journal_name

Pathology

journal_title

Pathology

authors

Tan YH,Liu Y,Eu KW,Ang PW,Li WQ,Salto-Tellez M,Iacopetta B,Soong R

doi

10.1080/00313020801911512

subject

Has Abstract

pub_date

2008-04-01 00:00:00

pages

295-8

issue

3

eissn

0031-3025

issn

1465-3931

pii

S0031-3025(16)32391-1

journal_volume

40

pub_type

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