Detection of BRAF V600E mutation by pyrosequencing

Y.H. Tan, Y. Liu, K.W. Eu, P.W. Ang, W.Q. Li, M. Salto-Tellez, Barry Iacopetta, R. Soong

Research output: Contribution to journalArticlepeer-review

78 Citations (Scopus)


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.
Original languageEnglish
Pages (from-to)295-298
Issue number3
Publication statusPublished - 2008


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