ARMS-qPCR Improves Detection Sensitivity of Earlier Diagnosis of Papillary Thyroid Cancers with Worse Prognosis Determined by BRAF V600E and TERT Promoter Co-existing Mutations.

Abstract

OBJECTIVE\nThe co-existence of BRAF V600E and TERT promoter mutation C228T/C250T has been extensively associated with thyroid cancer prognosis. Our study aimed to establish a sensitive method for mutation detection and to explore the correlation in detail.\n\n\nMETHODS\nBRAF and TERT promoter mutation status of 250 papillary thyroid cancer was determined using both Amplification Refractory Mutation System quantitative PCR (ARMS-qPCR) and Sanger sequencing to compare the sensitivity of these two methods. The associations between the mutation status and the clinicopathological features were then analyzed.\n\n\nRESULTS\nResults showed that ARMS-qPCR was more sensitive than Sanger (BRAF V600E: 75.2% (188/250) vs. 52.4% (131/250), p<0.001; TERT promoter C228T/C250T mutation: 12.0% (30/250) vs. 3.6% (9/250), p=0.001; Co-mutation (9.6% (24/250) vs. 3.2% (8/250), p=0.005). Both ARMS-qPCR and Sanger indicated that BRAF V600E and TERT promoter co-mutation patients had a higher diagnosis age, recurrence rate, and were related to a more advanced TNM stage and higher MACIS score. Moreover, ARMS-qPCR identified an earlier group stage which was younger, with a smaller tumor, and a lower recurrence rate compared with the BRAF V600E and TERT promoter co-existing mutation Sanger results. Besides, the newly identified group had a lower MACIS score and a lower TNM stage.\n\n\nCONCLUSION\nIn conclusion, patients with co-existing BRAF V600E and TERT promoter mutation have a worse prognosis. ARMS-qPCR, the more sensitive method, can be used to identify the early stages of patients with a potentially worse prognosis.