Five-Gene Testing Panel Refines Diagnostic Accuracy and Risk Stratification for Thyroid Nodules

A newly designed polymerase chain reaction panel targeting five specific genes provides refined diagnostic accuracy and risk stratification for thyroid nodules with indeterminate cytology. In an evaluation of fine-needle aspiration samples collected from 354 thyroid nodules, the panel demonstrated an overall diagnostic accuracy of 88.2 percent for cytologically indeterminate cases, accompanied by a sensitivity of 86.9 percent and a specificity of 95.0 percent.

Current clinical guidelines recommend incorporating molecular testing into the diagnostic evaluation of thyroid nodules when cytological examinations yield indeterminate results. However, established commercial testing kits rely on RNA-based methodologies that are expensive and remain largely inaccessible to patient populations outside of the United States. To address these limitations in accessibility and cost, researchers developed an alternative testing panel based on polymerase chain reaction technology to detect alterations across five specific genes: BRAF, HRAS, NRAS, TERT, and CCDC6-RET.

When fine-needle aspiration samples from the 354 thyroid nodules were evaluated using the five-gene panel, thyroid cancer-associated genomic alterations were detected in 205 nodules, corresponding to 57.9 percent of the total sample. Among the alterations identified, the BRAF V600E mutation was the most prevalent genomic event, occurring in 184 of the 354 evaluated nodules, or 52.0 percent of the cohort.

Clinical management and risk assessment pathways were documented following initial cytological evaluation. Irrespective of their molecular testing results, 160 nodules from 152 patients were directed to immediate surgical resection following fine-needle aspiration cytology. The remaining 194 nodules were categorized into three defined molecular risk tiers: low risk, encompassing nodules that were mutation-negative or harbored RAS mutations; intermediate risk, comprising nodules with BRAF mutations; and high risk, defined by the concurrent presence of BRAF or RAS mutations alongside TERT mutations.

Longitudinal clinical follow-up data gathered from the stratified cohort demonstrated that patients assigned to the intermediate-risk and high-risk tiers were significantly more likely to receive a confirmed diagnosis of thyroid cancer or experience disease recurrence compared to patients in the low-risk group. For thyroid nodules presenting with indeterminate cytology, the panel yielded a positive predictive value of 98.9 percent and a negative predictive value of 59.4 percent, indicating that the five-gene approach can deliver accessible and cost-effective molecular guidance to support clinical nodule management.

Subject of Research: Molecular testing panel for thyroid nodule diagnosis and risk stratification

Article Title: A five-gene molecular testing panel refines thyroid nodules diagnosis and risk stratification

Article References: Ying, Y., Zhou, J., Xie, Y., Liu, J., Ge, Y., Peng, Q., Zhu, Y., Liao, H., Li, Z., Dai, M., Luo, H., Wan, S., Li, X., & Zeng, X. (2026). A five-gene molecular testing panel refines thyroid nodules diagnosis and risk stratification. BMC Endocrine Disorders. https://doi.org/10.1186/s12902-026-02688-y

Image Credits: AI Generated

DOI: 10.1186/s12902-026-02688-y

Keywords: thyroid nodules, molecular testing, gene panel, indeterminate cytology, fine-needle aspiration, BRAF mutation, risk stratification, cancer diagnostics, five-gene, molecular, testing, panel