A mutation panel comprising BRAFV600E, NRASQ61R, and NRASQ61H replicated retrospective histopathological examination findings in differentiating benign goitre from malignant papillary thyroid cancer in a cohort of Malaysian patients.

Thyroid malignancy status is usually confirmed through histopathological examination (HPE) following thyroidectomy. In Malaysia, the application of molecular markers in pre-operative diagnosis of thyroid cancer remains unexplored. In this study, BRAF and NRAS gene mutation panel was assessed, and the results were compared with retrospective HPE findings. Malaysian patients with benign goitre (BTG: n=33) and papillary thyroid cancer (PTC: n=25; PTCa: n=20, PTCb: n=5) were recruited at Universiti Malaya Medical Centre from September 2019 to December 2022. PCR-direct DNA sequencing of BRAF^V600, NRAS^G12, NRAS^G13, and NRAS^Q61 was conducted on DNA extracted from the patients' thyroid tissue specimens following thyroidectomy and HPE. BRAF^V600E and NRAS^Q61R mutations showed absolute PTC-specificity with PTC-sensitivity of 32% and 28%, respectively. NRAS^Q61H demonstrated lower PTC-specificity (94%) but higher PTC-sensitivity (72%) compared to the BRAF^V600E and NRAS^Q61R mutations. Although the NRAS^G12 and NRAS^G13 variants were absent in this study, a novel NRAS^V14D mutation was detected in a PTCa patient. Unlike PTCb, coexistence of BRAF^V600E and NRAS^Q61 variants was commonly observed among the PTCa patients. Notably, all PTCb patients had NRAS^Q61H mutation with one patient carried both the NRAS^Q61H and BRAF^V600E mutations. Association analysis revealed potential link between gender, BRAF^V600E mutation and lymph node metastasis. In conclusion, mutation panel comprising BRAF^V600E, NRAS^Q61R, and NRAS^Q61H did not discriminate the two PTC subtypes but replicated the retrospective HPE findings in differentiating BTG from PTC. The application of this mutation panel in pre-operative diagnosis of thyroid nodules requires further validation in a larger sample size, preferably incorporating fineneedle aspirate biopsies.