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Published Online First: 13 February 2007. doi:10.1136/jcp.2006.040105
Journal of Clinical Pathology 2007;60:1211-1215
Copyright © 2007 by the BMJ Publishing Group Ltd & Association of Clinical Pathologists.

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*Thyroid Cancer

ORIGINAL ARTICLES

Detection of BRAF V600E activating mutation in papillary thyroid carcinoma using PCR with allele-specific fluorescent probe melting curve analysis

Leslie R Rowe1, Brandon G Bentz2, Joel S Bentz3

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

Correspondence to:
Dr Joel S Bentz, Department of Pathology, Rm 3860, Huntsman Cancer Hospital at the University of Utah Health Sciences Center, 1950 Circle of Hope, Salt Lake City, UT 84112, USA; joel.bentz{at}hsc.utah.edu

Background: A single hotspot mutation at nucleotide 1799 of the BRAF gene has been identified as the most common genetic event in papillary thyroid carcinoma (PTC), with a prevalence of 29–83%.

Aims: To use a PCR assay to molecularly characterise the BRAF activating point mutation in a series of PTC and benign thyroid cases and correlate the mutation results with histological findings.

Methods: Formalin-fixed paraffin-embedded (FFPE) sections were evaluated for the BRAF V600E mutation using LightCycler PCR with allele-specific fluorescent probe melting curve analysis (LCPCR).

Results: 42 (37 PTC; 5 benign) surgical tissue samples were analysed for the BRAF V600E activating point mutation. Using LCPCR and direct DNA sequencing, the BRAF mutation was identified in 23/37 (62.2%) PTC FFPE samples. DNA sequencing results demonstrated confirmation of the mutation.

Conclusions: Detection of BRAF-activating mutations in PTC suggests new approaches to management and treatment of this disease that may prove worthwhile. Identification of the BRAF V600E activating mutation in routine FFPE pathology samples by a rapid laboratory method such as LCPCR could have significant value.


Abbreviations: FFPE, formalin-fixed paraffin-embedded; FNA, fine needle aspirate; FVPTC, follicular variant of PTC; LCPCR, LightCycler PCR with allele-specific fluorescent probe melting curve analysis; MAPK, mitogen-activated protein kinase; PTC, papillary thyroid carcinoma; WT, wild type







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Copyright © 2007 by the BMJ Publishing Group Ltd & Association of Clinical Pathologists.