Peptide nucleic acid clamp polymerase chain reaction reveals a deletion mutation of the BRAF gene in papillary thyroid carcinoma: A case report
- Authors:
View Affiliations
Affiliations: Department of Laboratory Medicine and Genetics, Soonchunhyang University Bucheon Hospital and Soonchunhyang University College of Medicine, Bucheon, Gyeonggi 420-767, Republic of Korea
- Published online on: October 8, 2013 https://doi.org/10.3892/etm.2013.1332
-
Pages:
1550-1552
Metrics: Total
Views: 0 (Spandidos Publications: | PMC Statistics: )
Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )
This article is mentioned in:
Abstract
The BRAF point mutation is the most common genetic event in papillary thyroid carcinoma (PTC), occurring in 29-69% of such tumors. The V600E mutation accounts for up to 95% of all BRAF mutations. Therefore, the majority of diagnostic assays have been developed to detect only the V600E mutation of the BRAF gene. A peptide nucleic-acid (PNA)-clamp quantitative polymerase chain reaction (qPCR) was developed to detect the V600E mutation and other mutations in the BRAF gene. In this study, a 3-bp deletion mutation (c.1799_ 1801delTGA) was detected in a subject with a PTC by PNA clamp qPCR, in contrast with the results of allele‑specific (AS)-PCR. The mutant allele was not detected by AS-PCR, but was detected using PNA-clamp PCR. The atypical 3-bp deletion mutation (c.1799_1801delTGA) was identified by confirmatory PCR combined with sequencing. The conversion of codons 600 (GTG) and 601 (AAA) into a single codon (GAA) resulted in the insertion of a glutamic acid residue into the activation segment of the B-raf protein (p.V600_K601delinsE). In cases where PTC is highly suspected but no mutation is detected by AS-PCR specific for V600E, PNA clamp qPCR, which is complementary to other sequencing methods, should be performed in order to detect other mutations in the BRAF gene.
View References
1
|
Segev DL, Umbricht C and Zeiger MA:
Molecular pathogenesis of thyroid cancer. Surg Oncol. 12:69–90.
2003. View Article : Google Scholar : PubMed/NCBI
|
2
|
Davies H, Bignell GR, Cox C, et al:
Mutations of the BRAF gene in human cancer. Nature. 417:949–954.
2002. View Article : Google Scholar : PubMed/NCBI
|
3
|
Michaloglou C, Vredeveld LC, Mooi WJ and
Peeper DS: BRAF(E600) in benign and malignant human tumours.
Oncogene. 27:877–895. 2008. View Article : Google Scholar : PubMed/NCBI
|
4
|
Xing M: BRAF mutation in papillary thyroid
cancer: pathogenic role, molecular bases, and clinical
implications. Endocr Rev. 28:742–762. 2007. View Article : Google Scholar : PubMed/NCBI
|
5
|
Kim SK, Kim DL, Han HS, et al:
Pyrosequencing analysis for detection of a BRAFV600E mutation in an
FNAB specimen of thyroid nodules. Diagn Mol Pathol. 17:118–125.
2008. View Article : Google Scholar : PubMed/NCBI
|
6
|
Park SY, Park YJ, Lee YJ, et al: Analysis
of differential BRAF(V600E) mutational status in multifocal
papillary thyroid carcinoma: evidence of independent clonal origin
in distinct tumor foci. Cancer. 107:1831–1838. 2006. View Article : Google Scholar : PubMed/NCBI
|
7
|
Chung KW, Yang SK, Lee GK, et al:
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).
65:660–666. 2006. View Article : Google Scholar : PubMed/NCBI
|
8
|
Trovisco V, Soares P, Soares R, Magalhães
J, Sá-Couto P and Sobrinho-Simões M: A new BRAF gene mutation
detected in a case of a solid variant of papillary thyroid
carcinoma. Hum Pathol. 36:694–697. 2005. View Article : Google Scholar : PubMed/NCBI
|
9
|
Reifenberger J, Knobbe CB, Sterzinger AA,
et al: Frequent alterations of Ras signaling pathway genes in
sporadic malignant melanomas. Int J Cancer. 109:377–384. 2004.
View Article : Google Scholar : PubMed/NCBI
|
10
|
Chiosea S, Nikiforova M, Zuo H, et al: A
novel complex BRAF mutation detected in a solid variant of
papillary thyroid carcinoma. Endocr Pathol. 20:122–126. 2009.
View Article : Google Scholar : PubMed/NCBI
|
11
|
Jung CK, Im SY, Kang YJ, et al: Mutational
patterns and novel mutations of the BRAF gene in a large cohort of
Korean patients with papillary thyroid carcinoma. Thyroid.
22:791–797. 2012. View Article : Google Scholar : PubMed/NCBI
|
12
|
Milbury CA, Li J and Makrigiorgos GM:
PCR-based methods for the enrichment of minority alleles and
mutations. Clin Chem. 55:632–640. 2009. View Article : Google Scholar : PubMed/NCBI
|
13
|
Lee ST, Kim JY, Kown MJ, et al: Mutant
enrichment with 3′-modified oligonucleotides a practical PCR method
for detecting trace mutant DNAs. J Mol Diagn. 13:657–668. 2011.
|