Shifted Termination Assay (STA) Fragment Analysis to Detect BRAF V600 Mutations in Papillary Thyroid Carcinomas

Overview

Journal Diagn Pathol

Publisher Biomed Central

Specialty Pathology

Date 2013 Jul 26

PMID 23883275

Citations 4

Authors

So Young Kang

Soomin Ahn

Sun-Mi Lee

Ji Yun Jeong

Ji-Youn Sung

Young Lyun Oh

Kyoung-Mee Kim

Affiliations

Soon will be listed here.

Abstract

Background: BRAF mutation is an important diagnostic and prognostic marker in patients with papillary thyroid carcinoma (PTC). To be applicable in clinical laboratories with limited equipment, diverse testing methods are required to detect BRAF mutation.

Methods: A shifted termination assay (STA) fragment analysis was used to detect common V600 BRAF mutations in 159 PTCs with DNAs extracted from formalin-fixed paraffin-embedded tumor tissue. The results of STA fragment analysis were compared to those of direct sequencing. Serial dilutions of BRAF mutant cell line (SNU-790) were used to calculate limit of detection (LOD).

Results: BRAF mutations were detected in 119 (74.8%) PTCs by STA fragment analysis. In direct sequencing, BRAF mutations were observed in 118 (74.2%) cases. The results of STA fragment analysis had high correlation with those of direct sequencing (p < 0.00001, κ = 0.98). The LOD of STA fragment analysis and direct sequencing was 6% and 12.5%, respectively. In PTCs with pT3/T4 stages, BRAF mutation was observed in 83.8% of cases. In pT1/T2 carcinomas, BRAF mutation was detected in 65.9% and this difference was statistically significant (p = 0.007). Moreover, BRAF mutation was more frequent in PTCs with extrathyroidal invasion than tumors without extrathyroidal invasion (84.7% versus 62.2%, p = 0.001). To prepare and run the reactions, direct sequencing required 450 minutes while STA fragment analysis needed 290 minutes.

Conclusions: STA fragment analysis is a simple and sensitive method to detect BRAF V600 mutations in formalin-fixed paraffin-embedded clinical samples.

Virtual Slides: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/5684057089135749.

Citing Articles

The changing characteristics and molecular profiles of papillary thyroid carcinoma over time: a systematic review.

Vuong H, Altibi A, Abdelhamid A, Ngoc P, Quan V, Tantawi M Oncotarget. 2016; 8(6):10637-10649.

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A novel approach for the detection and genetic analysis of live melanoma circulating tumor cells.

Xu M, Cooke M, Steinmetz D, Karakousis G, Saxena D, Bartlett E PLoS One. 2015; 10(3):e0123376.

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Analysis of BRAF(V600E) mutation and DNA methylation improves the diagnostics of thyroid fine needle aspiration biopsies.

Zhang B, Liu S, Zhang Z, Wei J, Qu Y, Wu K Diagn Pathol. 2014; 9:45.

PMID: 24588959 PMC: 3944809. DOI: 10.1186/1746-1596-9-45.

The association between BRAF (V600E) mutation and pathological features in PTC.

Liu X, Yan K, Lin X, Zhao L, An W, Wang C Eur Arch Otorhinolaryngol. 2014; 271(11):3041-52.

PMID: 24389984 DOI: 10.1007/s00405-013-2872-7.

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