Detection of mutant BRAF alleles in the plasma of patients with metastatic melanoma Journal Article
| Authors: | Yancovitz, M.; Yoon, J.; Mikhail, M.; Gai, W.; Shapiro, R. L.; Berman, R. S.; Pavlick, A. C.; Chapman, P. B.; Osman, I.; Polsky, D. |
| Article Title: | Detection of mutant BRAF alleles in the plasma of patients with metastatic melanoma |
| Abstract: | Mutations in the BRAF oncogene at amino acid 600 have been reported in 40 to 70% of human metastatic melanoma tissues, and the critical role of BRAF in the biology of melanoma has been established. Sampling the blood compartment to detect the mutational status of a solid tumor represents a highly innovative advance in cancer medicine, and such an approach could have advantages over tissue-based techniques. We report the development of a fluorescence-based polymerase chain reaction (PCR) assay to detect mutant BRAF alleles in plasma. A mutant-specific PCR assay was optimized to specifically amplify the mutant BRAF allele without amplifying the wild-type allele. Experiments mixing DNA from a BRAF mutant melanoma cell line with wild-type human placental DNA in varying proportions were performed to determine the threshold of this assay and to compare it with routine DNA sequencing. The assay was then applied to tissue and plasma specimens from patients with metastatic melanoma. The assay detected 0.1 ng of mutant DNA mixed in 100 ng of wild-type DNA and was 500-fold more sensitive than DNA sequencing. The assay detected mutant BRAF alleles in plasma samples from 14 of 26 (54%) metastatic melanoma patients. These data demonstrate the feasibility of blood-based testing for BRAF mutations in metastatic melanoma patients. Copyright © American Society for Investigative Pathology and the Association for Molecular Pathology. |
| Keywords: | adult; clinical article; controlled study; human tissue; aged; aged, 80 and over; middle aged; unclassified drug; gene mutation; human cell; genetics; molecular genetics; sensitivity and specificity; mutant protein; polymerase chain reaction; sensitivity analysis; protein blood level; allele; melanoma; metastasis; alleles; cancer cell culture; cell line, tumor; wild type; blood; feasibility study; dna; molecular sequence data; blood analysis; nucleotide sequence; tumor cell line; dna, neoplasm; neoplasm metastasis; base sequence; temperature; dna sequence; metastasis potential; dna mutational analysis; mutant proteins; genetic screening; fluorescence analysis; b raf kinase; placenta; proto-oncogene proteins b-raf; braf protein, human |
| Journal Title: | Journal of Molecular Diagnostics |
| Volume: | 9 |
| Issue: | 2 |
| ISSN: | 1525-1578 |
| Publisher: | Elsevier Science, Inc. |
| Date Published: | 2007-04-01 |
| Start Page: | 178 |
| End Page: | 183 |
| Language: | English |
| DOI: | 10.2353/jmoldx.2007.060135 |
| PUBMED: | 17384209 |
| PROVIDER: | scopus |
| PMCID: | PMC1867452 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 11" - "Export Date: 17 November 2011" - "CODEN: JMDIF" - "Source: Scopus" |
