Comparison of Three BRAF Mutation Tests in Formalin-Fixed Paraffin Embedded Clinical Samples
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Background: Recently, BRAF inhibitors showed dramatic treatment outcomes in BRAF V600 mutant melanoma. Therefore, the accuracy of BRAF mutation test is critical. Methods: BRAF mutations were tested by dual-priming oligonucleotide-polymerase chain reaction (DPO-PCR), direct sequencing and subsequently retested with a real-time PCR assay, cobas 4800 V600 mutation test. In total, 64 tumors including 34 malignant melanomas and 16 papillary thyroid carcinomas were analyzed. DNA was extracted from formalin-fixed paraffin embedded tissue samples and the results of cobas test were directly compared with those of DPO-PCR and direct sequencing. Results: BRAF mutations were found in 23 of 64 (35.9%) tumors. There was 9.4% discordance among 3 methods. Out of 6 discordant cases, 4 cases were melanomas; 3 cases were BRAF V600E detected only by cobas test, but were not detected by DPO-PCR and direct sequencing. One melanoma patient with BRAF mutation detected only by cobas test has been on vemurafenib treatment for 6 months and showed a dramatic response to vemurafenib. DPO-PCR failed to detect V600K mutation in one case identified by both direct sequencing and cobas test. Conclusions: In direct comparison of the currently available DPO-PCR, direct sequencing and realtime cobas test for BRAF mutation, real-time PCR assay is the most sensitive method.
Abstract
Background: Recently, BRAF inhibitors showed dramatic treatment outcomes in BRAF V600 mutant melanoma. Therefore, the accuracy of BRAF mutation test is critical. Methods: BRAF mutations were tested by dual-priming oligonucleotide-polymerase chain reaction (DPO-PCR), direct sequencing and subsequently retested with a real-time PCR assay, cobas 4800 V600 mutation test. In total, 64 tumors including 34 malignant melanomas and 16 papillary thyroid carcinomas were analyzed. DNA was extracted from formalin-fixed paraffin embedded tissue samples and the results of cobas test were directly compared with those of DPO-PCR and direct sequencing. Results: BRAF mutations were found in 23 of 64 (35.9%) tumors. There was 9.4% discordance among 3 methods. Out of 6 discordant cases, 4 cases were melanomas; 3 cases were BRAF V600E detected only by cobas test, but were not detected by DPO-PCR and direct sequencing. One melanoma patient with BRAF mutation detected only by cobas test has been on vemurafenib treatment for 6 months and showed a dramatic response to vemurafenib. DPO-PCR failed to detect V600K mutation in one case identified by both direct sequencing and cobas test. Conclusions: In direct comparison of the currently available DPO-PCR, direct sequencing and realtime cobas test for BRAF mutation, real-time PCR assay is the most sensitive method.
참고문헌 (22)
- Gray-Schopfer V, "Melanoma biology and new targeted therapy", Nature, 445, pp.851~857 (2007)
- Cummins DL, "Cutaneous malignant melanoma", Mayo Clin Proc, 81, pp.500~507 (2006)
- Cohen C, "Mitogen-actived protein kinase activation is an early event in melanoma progression", Clin Cancer Res, 8, pp.3728~3733 (2002)
- Platz A, "Human cutaneous melanoma; a review of NRAS and BRAF mutation frequencies in relation to histogenetic subclass and body site", Mol Oncol, 1, pp.395~405 (2008)
- Long GV, "Prognostic and clinicopathologic associations of oncogenic BRAF in metastatic melanoma", J Clin Oncol, 29, pp.1239~1246 (2011)
- Chapman PB, "Improved survival with vemurafenib in melanoma with BRAF V600E mutation", N Engl J Med, 364, pp.2507~2516 (2011)
- Sosman JA, "Survival in BRAF V600-mutant advanced melanoma treated with vemurafenib", N Engl J Med, 366, pp.707~714 (2012)
- Hatzivassiliou G, "RAF inhibitors prime wild-type RAF to activate the MAPK pathway and enhance growth", Nature, 464, pp.431~435 (2010)
- Halaban R, "PLX4032, a selective BRAF(V600E) kinase inhibitor, activates the ERK pathway and enhances cell migration and proliferation of BRAF melanoma cells", Pigment Cell Melanoma Res, 23, pp.190~200 (2010)
- Poulikakos PI, "RAF inhibitors transactivate RAF dimers and ERK signalling in cells with wild-type BRAF", Nature, 464, pp.427~430 (2010)
- Anderson S, "Multisite analytic performance studies of a real-time polymerase chain reaction assay for the detection of BRAF V600E mutations in formalin-fixed, paraffin-embedded tissue specimens of malignant melanoma", Arch Pathol Lab Med, 136, pp.1385~1391 (2012)
- Halait H, "Analytical performance of a real-time PCR-based assay for V600 mutations in the BRAF gene, used as the companion diagnostic test for the novel BRAF inhibitor vemurafenib in metastatic melanoma", Diagn Mol Pathol, 21, pp.1~8 (2012)
- Bloom KJ, "Molecular testing for BRAF V600 mutations in the BRIM-2 trial of the BRAF inhibitor vemurafenib in metastatic melanoma", J Clin Oncol, 29, pp.10523~ (2011)
- Dong SM, "Progressive methylation during the serrated neoplasia pathway of the colorectum", Mod Pathol, 18, pp.170~178 (2005)
- Woodman SE, "New strategies in melanoma: molecular testing in advanced disease", Clin Cancer Res, 18, pp.1195~1200 (2012)
- Tan YH, "Detection of BRAF V600E mutation by pyrosequencing", Pathology, 40, pp.295~298 (2008)
- Rueschoff J, "Molecular testing for BRAF V600 mutations in the phase III trial of the selective BRAF inhibitor vemurafenib in metastatic melanoma: a comparison of the cobas 4800 BRAF V600E mutation test and Sanger sequencing", Pigment Cell Melanoma Res, 24, pp.1023~ (2011)
- Lee HJ, "Detection of BRAF mutations in thyroid nodules by allele-specific PCR using a dual priming oligonucleotide system", Am J Clin Pathol, 133, pp.802~808 (2010)
- Chung KW, "Detection of BRAFV600E mutation on fine needle aspiration specimens of thyroid nodule refines cyto-pathology diagnosis, especially in BRAF600E mutation-prevalent area", Clin Endocrinol (Oxf), 65, pp.660~666 (2006)
- Lin J, "Polyclonality of BRAF mutations in primary melanoma and the selection of mutant alleles during progression", Br J Cancer, 104, pp.464~468 (2011)
- Sullivan RJ, "BRAF in melanoma: pathogenesis, diagnosis, inhibition, and resistance", J Skin Cancer, 2011, pp.423239~ (2011)
- Si L, "Prevalence of BRAF V600E mutation in Chinese melanoma patients: large scale analysis of BRAF and NRAS mutations in a 432-case cohort", Eur J Cancer, 48, pp.94~100 (2012)