Simultaneously Detection of 50 Mutations at 20 Sites in the BRAF and RAS Genes by Multiplexed Single-Nucleotide Primer Extension Assay Using Fine-Needle Aspirates of Thyroid Nodules

Aaron A Stence; Michael P Gailey; Robert A Robinson; Chris S Jensen; Deqin Ma

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Simultaneously Detection of 50 Mutations at 20 Sites in the BRAF and RAS Genes by Multiplexed Single-Nucleotide Primer Extension Assay Using Fine-Needle Aspirates of Thyroid Nodules

Journal article

Peer reviewed

Simultaneously Detection of 50 Mutations at 20 Sites in the BRAF and RAS Genes by Multiplexed Single-Nucleotide Primer Extension Assay Using Fine-Needle Aspirates of Thyroid Nodules

Aaron A Stence, Michael P Gailey, Robert A Robinson, Chris S Jensen and Deqin Ma

The Yale journal of biology & medicine, Vol.88(4), pp.351-358

12/2015

PMCID: PMC4654183

PMID: 26604858

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Abstract

Fine-needle aspiration (FNA) is commonly used for primary evaluation of thyroid nodules. Twenty to 30 percent of thyroid nodules remain indeterminate after FNA evaluation. Studies show the BRAF p.V600E to be highly specific for papillary thyroid carcinoma (PTC), while RAS mutations carry up to 88 percent positive predictive value for malignancy. We developed a two-tube multiplexed PCR assay followed by single-nucleotide primer extension assay for simultaneous detection of 50 mutations in the BRAF (p.V600E, p.K601E/Q) and RAS genes (KRAS and NRAS codons 12, 13, 19, 61 and HRAS 61) using FNA smears of thyroid nodules. Forty-two FNAs and 27 paired formalin-fixed, paraffin-embedded (FFPE) tissues were tested. All BRAF p.V600E-positive FNA smears (five) carried a final diagnosis of PTC on resection. RAS mutations were found in benign as well as malignant lesions. Ninety-two percent concordance was observed between FNA and FFPE tissues. In conclusion, our assay is sensitive and reliable for simultaneous detection of multiple BRAF/RAS mutations in FNA smears of thyroid nodules.

Biopsy, Fine-Needle

Thyroid Nodule - genetics

Genetic Predisposition to Disease - genetics

Multiplex Polymerase Chain Reaction - methods

ras Guanine Nucleotide Exchange Factors - genetics

Proto-Oncogene Proteins B-raf - genetics

Thyroid Gland - pathology

Thyroid Nodule - pathology

Humans

Thyroid Nodule - diagnosis

Polymorphism, Single Nucleotide - genetics

Details

Title: Subtitle: Simultaneously Detection of 50 Mutations at 20 Sites in the BRAF and RAS Genes by Multiplexed Single-Nucleotide Primer Extension Assay Using Fine-Needle Aspirates of Thyroid Nodules
Creators: Aaron A Stence - Department of Pathology, University of Iowa Hospitals and Clinics, Iowa City, Iowa
Michael P Gailey - Department of Pathology, University of Iowa Hospitals and Clinics, Iowa City, Iowa
Robert A Robinson - Department of Pathology, University of Iowa Hospitals and Clinics, Iowa City, Iowa
Chris S Jensen - Department of Pathology, University of Iowa Hospitals and Clinics, Iowa City, Iowa
Deqin Ma - Department of Pathology, University of Iowa Hospitals and Clinics, Iowa City, Iowa
Resource Type: Journal article
Publication Details: The Yale journal of biology & medicine, Vol.88(4), pp.351-358
Publisher: United States
PMID: 26604858
PMCID: PMC4654183
ISSN: 0044-0086
eISSN: 1551-4056
Language: English
Date published: 12/2015
Academic Unit: Oral Pathology, Radiology and Medicine; Pathology
Record Identifier: 9984047794702771

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