التفاصيل البيبلوغرافية
| العنوان: |
Detection of Mutant BRAFAlleles in the Plasma of Patients with Metastatic Melanoma |
| المؤلفون: |
Yancovitz, Molly, Yoon, Joanne, Mikhail, Maryann, Gai, Weiming, Shapiro, Richard L., Berman, Russell S., Pavlick, Anna C., Chapman, Paul B., Osman, Iman, Polsky, David |
| المصدر: |
The Journal of Molecular Diagnostics; April 2007, Vol. 9 Issue: 2 p178-183, 6p |
| مستخلص: |
Mutations in the BRAFoncogene at amino acid 600 have been reported in 40 to 70% of human metastatic melanoma tissues, and the critical role of BRAFin 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 BRAFalleles in plasma. A mutant-specific PCR assay was optimized to specifically amplify the mutant BRAFallele without amplifying the wild-type allele. Experiments mixing DNA from a BRAFmutant 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 BRAFalleles in plasma samples from 14 of 26 (54%) metastatic melanoma patients. These data demonstrate the feasibility of blood-based testing for BRAFmutations in metastatic melanoma patients. |
| قاعدة البيانات: |
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