Comparative analysis of IHC, AMRS-PCR, and ddPCR for BRAF V600E mutation detection in papillary thyroid carcinoma
Year: 2025
Type: article
Study design: Observational Study
Abstract: • AMRS-PCR demonstrates superior sensitivity and specificity for BRAF V600E mutation detection in papillary thyroid carcinoma compared to IHC. • ddPCR effectively verifies low-frequency BRAF V600E mutations, resolving discordant IHC and AMRS-PCR results and highlighting its role as a confirmatory method. • A multimodal diagnostic approach, integrating AMRS-PCR for routine screening and ddPCR for ambiguous cases, enhances the accuracy of BRAF V600E mutation detection. • Accurate BRAF V600E detection with advanced molecular methods is crucial for precise risk stratification and individualized therapeutic decisions in papillary thyroid carcinoma. The BRAF V600E mutation is the most frequent driver alteration in papillary thyroid carcinoma (PTC) and plays an important role in diagnosis, risk stratification, and treatment planning. Commonly used detection methods include immunohistochemistry (IHC), amplification-refractory mutation system PCR (AMRS-PCR), and droplet digital PCR (ddPCR). However, their relative performance and their ability to identify low-frequency mutations have not been fully clarified. This retrospective study included 1,571 patients diagnosed with PTC between December 2021 and January 2025. All patients underwent IHC testing, and 871 also received AMRS-PCR for BRAF V600E detection. Sixty cases with discordant IHC and AMRS-PCR results were further analyzed using ddPCR. Detection rates, concordance, sensitivity, specificity, and variant allele frequency (VAF) were compared across the three methods. Among the 871 patients with both IHC and AMRS-PCR results, the overall concordance rate was 93.2%, and 59 cases showed discordant findings. ddPCR confirmed the presence of BRAF V600E in 24 discordant cases (40.7%). The ddPCR positivity rate was higher in the IHC-negative and AMRS-PCR-positive group than in the IHC-positive and AMRS-PCR-negative group (52.2% vs. 33.3%, P = 0.033), and the former group also showed a markedly higher VAF (mean 6.97% vs. 2.82%). AMRS-PCR demonstrated better sensitivity and specificity (75.0% and 85.7%) than IHC (25.0% and 14.3%). AMRS-PCR provides greater sensitivity and specificity than IHC for detecting BRAF V600E mutations and is appropriate for routine molecular testing. ddPCR offers excellent sensitivity for low-frequency mutations and is valuable for resolving discordant results. A multimodal testing strategy may improve detection accuracy and support personalized management of patients with PTC.
Institution: Shaanxi Provincial People's Hospital
Language: English
Subfield: Endocrinology, Diabetes and Metabolism
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $2,740
Award: G425512669
OpenAlex ID: W7117353539