Comparison of Three BRAF Mutation Tests in Formalin-Fixed Paraffin Embedded Clinical Samples.
- Author:
Soomin AHN
1
;
Jeeyun LEE
;
Ji Youn SUNG
;
So Young KANG
;
Sang Yun HA
;
Kee Taek JANG
;
Yoon La CHOI
;
Jung Sun KIM
;
Young Lyun OH
;
Kyoung Mee KIM
Author Information
1. Department of Pathology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea. kkmkys@skku.edu
- Publication Type:Original Article
- Keywords:
BRAF mutation;
Melanoma;
Real-time polymerase chain reaction;
Sanger sequencing;
Dual-priming oligonucleotide-PCR
- MeSH:
DNA;
Humans;
Indoles;
Melanoma;
Paraffin;
Real-Time Polymerase Chain Reaction;
Sulfonamides;
Thyroid Neoplasms
- From:Korean Journal of Pathology
2013;47(4):348-354
- CountryRepublic of Korea
- Language:English
-
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 real-time cobas test for BRAF mutation, real-time PCR assay is the most sensitive method.
