Detection of BRAF V600E mutation by pyrosequencing

doi:10.1080/00313020801911512

Pathology

Volume 40, Issue 3, April 2008, Pages 295-298

https://doi.org/10.1080/00313020801911512 Get rights and content

Summary

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.

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View full text

Detection of BRAF V600E mutation by pyrosequencing

Summary

Introduction

Methods

Results

Conclusions

Mutat Res

J Mol Diagn

J Mol Diagn

Mutations of the BRAF gene in human cancer

Nature

BRAF mutation is frequently present in sporadic colorectal cancer with methylated hMLH1, but not in hereditary nonpolyposis colorectal cancer

Clin Cancer Res