摘要： Background Somatic BRAF mutation is frequently observed in papillary thyroid carcinoma (PTC). Recent evidence suggests that PTCs are heterogeneous tumors containing a subclonal or oligoclonal occurrence of BRAF mutation. Conflicting results have been reported concerning the prognostic significance of the mutant allele frequency. Our present aim was to investigate the association between the percentage of BRAF c.1799T > A (p.Val600Glu) alleles and clinicopathological parameters in PTC. Methods Genomic DNA was extracted from fresh-frozen specimens obtained from 50 PTC patients undergoing total thyroidectomy. The BRAF mutation status was determined by Sanger sequencing. The percentage of mutant BRAF alleles was quantified by mass spectrometric genotyping, pyrosequencing, and competitive allele-specific TaqMan PCR (castPCR). Results Positive rate of BRAF mutation was 72 % by Sanger sequencing, 82 % by mass spectrometric genotying, and 84 % by pyrosequencing or castPCR. The average percentage of mutant BRAF alleles was 22.5, 31, and 30.7 %, respectively. There was a good correlation among three quantification methods (Spearman’s rho = 0.87–0.97; p < 0.0001). The mutant allele frequency was significantly correlated with tumor size (rho = 0.47–0.52; p < 0.01) and extrathyroidal invasion. The frequency showed no difference in pathological lymph node metastasis. Conclusions The percentage of mutant BRAF alleles is positively associated with tumor burden and extrathyroidal invasion in PTC. Relatively good correlations exist among mass spectrometric genotyping, pyrosequencing, and castPCR in quantification of mutant BRAF allele frequency. 其他題名： Ann Surg Oncol 出版者： Boston: Springer Science and Business Media LLC 出版日期： 2014-12-01 出處： Annals of Surgical Oncology, 2014-12, Vol.21 (S4), p.619-626 版權： Society of Surgical Oncology 2014 識別號： ISSN: 1068-9265 識別號： ISSN: 1534-4681 識別號： EISSN: 1534-4681 識別號： DOI: 10.1245/s10434-014-3723-5 識別號： PMID: 24748129
