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Detection of BRAF V600E mutation with thyroid tissue using pyrosequencing: comparison with PNA-clamping and real-time PCR.

Authors
 Seong-Ho Kang  ;  Ju Yeon Pyo  ;  Seok-Woo Yang  ;  Soon Won Hong 
Citation
 AMERICAN JOURNAL OF CLINICAL PATHOLOGY, Vol.139(6) : 759-764, 2013 
Journal Title
AMERICAN JOURNAL OF CLINICAL PATHOLOGY
ISSN
 0002-9173 
Issue Date
2013
MeSH
Adolescent ; Adult ; Aged ; Carcinoma/diagnosis* ; Carcinoma/genetics ; Carcinoma, Papillary ; Humans ; Middle Aged ; Mutation ; Peptide Nucleic Acids ; Polymerase Chain Reaction/methods ; Prognosis ; Proto-Oncogene Proteins B-raf/genetics* ; Real-Time Polymerase Chain Reaction/methods ; Sequence Analysis, DNA* ; Thyroid Gland/pathology ; Thyroid Neoplasms/diagnosis* ; Thyroid Neoplasms/genetics
Keywords
BRAF ; Peptide nucleic acid ; Pyrosequencing ; Quantification ; Sensitivity
Abstract
We used pyrosequencing, peptide nucleic acid (PNA)–clamping polymerase chain reaction (PCR), and real-time PCR to detect the BRAF V600E mutation and to investigate the prognostic effect of the BRAF V600E mutation in paraffin block specimens from 100 patients diagnosed with papillary thyroid carcinoma. Positive rates of PNA-clamping PCR, real-time PCR, and pyrosequencing were 66%, 70%, and 68%, respectively. Pyrosequencing and PNA-clamping PCR detected mutant type in a 99:1 (wild-type: mutant) DNA concentration, and PNA-clamping PCR detected mutant type in a 99.5:0.5 DNA concentration. Clamping PCR showed higher κ value than real-time PCR (0.729 vs 0.626). The BRAF V600E mutation was associated with an advanced stage of cancer (P = .045) and was found to be associated with poor prognostic factors. This study suggests that pyrosequencing can be as sensitive as real-time PCR and that PNA-clamping PCR is a sensitive and reliable method to detect the BRAF V600E mutation
Full Text
http://ajcp.ascpjournals.org/content/139/6/759.long
DOI
10.1309/AJCPN3ULH6YWBHPH
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Pathology (병리학교실) > 1. Journal Papers
Yonsei Authors
Yang, Seok Woo(양석우)
Hong, Soon Won(홍순원) ORCID logo https://orcid.org/0000-0002-0324-2414
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/87307
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