Rapid screening of 4000 individuals for germ-line variations in the BRAF gene

James, Michael R., Dumeni, Troy, Stark, Mitchell S., Duffy, David L., Montgomery, Grant W., Martin, Nicholas G. and Hayward, Nicholas K. (2006) Rapid screening of 4000 individuals for germ-line variations in the BRAF gene. Clinical Chemistry, 52 9: 1675-1678. doi:10.1373/clinchem.2006.070169

Author James, Michael R.
Dumeni, Troy
Stark, Mitchell S.
Duffy, David L.
Montgomery, Grant W.
Martin, Nicholas G.
Hayward, Nicholas K.
Title Rapid screening of 4000 individuals for germ-line variations in the BRAF gene
Journal name Clinical Chemistry   Check publisher's open access policy
ISSN 0009-9147
Publication date 2006
Sub-type Article (original research)
DOI 10.1373/clinchem.2006.070169
Volume 52
Issue 9
Start page 1675
End page 1678
Total pages 4
Editor D. E. Bruns
Place of publication Washington, U.S.A.
Publisher American Association for Clinical Chemistry
Collection year 2006
Language eng
Subject C1
321011 Medical Genetics
730107 Inherited diseases (incl. gene therapy)
Abstract Background: The BRAF gene is frequently somatically altered in malignant melanoma. A majority of variations are at the valine 600 residue leading to a V600E substitution that constitutively activates the kinase. We screened 4000 patient and control DNAs for germ-line variations at the valine 600 residue. Methods: We developed a novel assay by adapting single-base variation assays and software for MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry to screen for all 5 reported variants at codon 600 of the BRAF gene. We screened a case-control collection comprising samples from 1082 melanoma patients and 154 of their unaffected relatives from 1278 families and from 2744 individuals from 659 unselected twin families with no history of melanoma. A panel of 66 melanoma cell lines was used for variation-positive controls. Results: All melanoma cell lines that we had found previously to carry a codon 600 variation were verified in this study. Three of the 4 possible variants (V600E n = 47, V600K n = 2, V600R n = 1) were detected, but no case of V600D was available. No germ-line variants were found in the samples from the 3980 melanoma patients or from the control individuals. Conclusions: This new assay is a high-throughput, automated alternative to standard sequencing and can be used as a rapid initial screen for somatic variants associated with melanoma. Germ-line variants at valine 600 are unlikely to exist and do not contribute to the reported role of the BRAF gene in melanoma predisposition. (c) 2006 American Association for Clinical Chemistry.
Keyword Medical Laboratory Technology
Melanoma Susceptibility
Melanocytic Neoplasia
Metastatic Melanoma
Oncogenic Mutations
Signaling Pathway
Ras Mutations
Q-Index Code C1
Institutional Status Non-UQ

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Created: Wed, 15 Aug 2007, 09:50:50 EST