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 RESEARCH
CytoJournal 2006,  3:10

Utility of BRAF V600E mutation detection in cytologically indeterminate thyroid nodules


1 Institute for Clinical and Experimental Pathology, Associated Regional and University Pathologists (ARUP) Laboratories, Salt Lake City, UT, USA
2 Division of Otolaryngology, Department of Surgery, University of Utah, Salt Lake City, UT, USA
3 Institute for Clinical and Experimental Pathology, Associated Regional and University Pathologists (ARUP) Laboratories; Department of Pathology, University of Utah, Salt Lake City, UT, USA

Correspondence Address:
Joel S Bentz
Institute for Clinical and Experimental Pathology, Associated Regional and University Pathologists (ARUP) Laboratories; Department of Pathology, University of Utah, Salt Lake City, UT
USA
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Source of Support: None, Conflict of Interest: None


DOI: 10.1186/1742-6413-3-10

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Background: Fine needle aspiration (FNA) is widely utilized for evaluation of patients with thyroid nodules. However, approximately 30% are indeterminate for malignancy. Recently, a mutation in the BRAF gene has been reported to be the most common genetic event in papillary thyroid carcinoma (PTC). In this retrospective study, we assessed the utility of BRAF V600E mutation detection for refining indeterminate preoperative cytologic diagnoses in patients with PTC. Methods: Archival indeterminate thyroid FNAs and corresponding formalin-fixed, paraffin-embedded (FFPE) surgical samples with PTC were identified in our patient files. DNA extracted from slide scape lysates and 5 μm FFPE sections were evaluated for the BRAF V600E mutation using LightCycler PCR and fluorescent melting curve analysis (LCPCR). Amplification products that showed deviation from the wild-type genomic DNA melting peak, discordant FNA and FFPE matched pairs, and all benign control samples, underwent direct DNA sequencing. Results: A total of 19 indeterminate thyroid FNAs demonstrating PTC on FFPE surgical samples were included in the study. Using BRAF mutation analysis, the preoperative diagnosis of PTC was confirmed in 3/19 (15.8%) FNA samples that could not be conclusively diagnosed on cytology alone. However, 9/19 (47.4%) FFPE tissue samples were positive for the V600E mutation. Of the discordant pairs, 5/6 FNAs contained less than 50% tumor cells. Conclusion: When used with indeterminate FNA samples, BRAF mutation analysis may be a useful adjunct technique for confirming the diagnosis of malignancy in an otherwise equivocal case. However, overall tumor cell content of some archival FNA smear slides is a limiting factor for mutation detection.






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