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.