TEST:

Afirma® Genomic Sequencing Classifier

NRAS p.Q61R/K comprises 99% of reported NRAS variants. TERTp is the most frequent co-mutation. Among NRAS p.Q61R/K nodules, there are 2 different clusters of samples based on the activity of hallmarks of cancer pathways. Further studies correlating these clusters with clinical and pathological outcomes are necessary.

P=N/A, N=50, Recruiting, Columbia University | Trial completion date: Aug 2027 --> Aug 2028 | Trial primary completion date: Aug 2026 --> Aug 2027

The frequency of PLEKHS1p mutations did not strongly correlate with the severity of thyroid cancer but the surgical sample size was limited. Further research is needed to clarify the role of PLEKHS1p mutations in thyroid nodules and cancer.

In young patients with ITN, the Afirma GSC demonstrated an excellent negative predictive value when defining a TN result by histology or clinical follow-up.

Afirma GSC has a high BCR and improved performance over earlier generation molecular tests in oncocytic thyroid nodules, particularly for Bethesda IV. However, the PPV in Bethesda III nodules remains low, especially in the presence of Hashimoto's thyroiditis.

A GSC-suspicious result combined with TIRADS 5 significantly improved the positive predictive value, thereby enhancing risk stratification in AUS thyroid nodules, while other diagnostic parameters remained unchanged.

P=N/A, N=328, Active, not recruiting, Jonsson Comprehensive Cancer Center | Trial completion date: Dec 2026 --> Dec 2027 | Trial primary completion date: Dec 2025 --> Dec 2026

Both molecular tests demonstrate high NPV but low specificity; neither is clearly superior. Future research should prioritise randomised controlled trials, long-term follow-up of unoperated nodules, and direct comparisons of molecular tests.

These guidelines emphasize the need for standardized protocols and equitable access to such testing. Molecular diagnostics should be embraced as complementary tools within multidisciplinary care to optimize patient outcomes while reducing unnecessary interventions in thyroid nodule management.

AUS-Nuclear carries a substantially higher ROM than AUS-Other, with a ROM (54.7%) comparable to the reported positive predictive values of molecular assays such as Afirma GSC (47%, 95% CI: 36%-58%) and ThyroSeq v3 (66%, 95% CI: 56%-75%). These findings support the clinical utility of the two-tiered AUS subclassification in enhancing risk stratification, particularly in settings where molecular testing is not routinely available.