A multisite blinded study for the detection of BRAF mutations in formalin-fixed, paraffin-embedded malignant melanoma.

Abstract:

:Melanoma patients with BRAF mutations respond to treatment with vemurafenib, thus creating a need for accurate testing of BRAF mutation status. We carried out a blinded study to evaluate various BRAF mutation testing methodologies in the clinical setting. Formalin-fixed, paraffin-embedded melanoma samples were macrodissected before screening for mutations using Sanger sequencing, single-strand conformation analysis (SSCA), high resolution melting analysis (HRM) and competitive allele-specific TaqMan® PCR (CAST-PCR). Concordance of 100% was observed between the Sanger sequencing, SSCA and HRM techniques. CAST-PCR gave rapid and accurate results for the common V600E and V600K mutations, however additional assays are required to detect rarer BRAF mutation types found in 3-4% of melanomas. HRM and SSCA followed by Sanger sequencing are effective two-step strategies for the detection of BRAF mutations in the clinical setting. CAST-PCR was useful for samples with low tumour purity and may also be a cost-effective and robust method for routine diagnostics.

journal_name

Sci Rep

journal_title

Scientific reports

authors

Richter A,Grieu F,Carrello A,Amanuel B,Namdarian K,Rynska A,Lucas A,Michael V,Bell A,Fox SB,Hewitt CA,Do H,McArthur GA,Wong SQ,Dobrovic A,Iacopetta B

doi

10.1038/srep01659

subject

Has Abstract

pub_date

2013-01-01 00:00:00

pages

1659

issn

2045-2322

pii

srep01659

journal_volume

3

pub_type

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