ARMS-qPCR-Based Detection of BRAF and TERT Promoter Mutations: A Cost-Effective Strategy for Molecular Diagnosis of Papillary Thyroid Carcinoma.

This study aimed to establish a rapid and sensitive method for detecting BRAF V600E, TERT C228T, and TERT C250T mutations using amplification-refractory mutation system-quantitative PCR (ARMS-qPCR) combined with TaqMan probes. We systematically evaluated the assay's accuracy, sensitivity, repeatability, and specificity in the laboratory and assessed detection performance in clinical samples, in which a total of 40 formalin-fixed paraffin-embedded samples (34 papillary thyroid carcinoma [PTC] tissues and 6 normal tissues) were analyzed by ARMS-qPCR, commercial kits, and Sanger sequencing. In the laboratory, the ARMS-qPCR assay exhibited superior performance of accuracy, sensitivity, repeatability, and specificity. In clinical samples, ARMS-qPCR detected BRAF V600E mutation in 82.35% (28/34) and TERT C228T in 5.9% (2/34), which the commercial kit performed equally, while Sanger sequencing identified BRAF V600E in 50% (17/34) and TERT C228T in 2.9% (1/34). Simultaneously, neither method detected TERT C250T mutations. The ARMS-qPCR method demonstrated significantly higher sensitivity than Sanger sequencing, along with advantages of rapid turnaround (2-3 h), cost-effectiveness, and broad applicability. These findings highlight ARMS-qPCR as a robust, sensitive, and efficient approach for detecting BRAF and TERT promoter mutations in PTCs, offering substantial potential for clinical molecular diagnostics and personalized management of thyroid cancer.