Assessing the clinical diagnostic utility of multiplex ddPCR assays in thyroid nodules.
Six-gene ddPCR panel plus cytology greatly improved thyroid nodule diagnostic accuracy.
Assessing the clinical diagnostic utility of multiplex ddPCR assays in thyroid nodules.
Ultrasound-guided fine-needle aspiration cytology (fine-needle aspiration cytology) is the standard method for evaluating thyroid nodules, but 20–40% of specimens are still cytologically indeterminate (Bethesda III–IV), leading to repeat biopsies or diagnostic lobectomy.
We hypothesized that a multiplex droplet digital PCR (ddPCR) assay covering key thyroid-cancer driver mutations would increase the pre-operative diagnostic yield.
The assays were subsequently applied to 210 prospectively collected fine-needle aspiration (fine-needle aspiration) specimens from 201 patients, with parallel ARMS-PCR testing for BRAF p.
combining ddPCR with cytology matched surgical diagnosis far more often than cytology alone
The six-gene ddPCR panel demonstrated 100% analytical specificity and a limit of detection of 0.1% variant allele frequency (variant allele frequency).
In clinical samples, 103 of 210 nodules (49%) harbored at least one mutation, with BRAF p.
V600E being the most prevalent alteration (81/103, 78.6%). ddPCR identified five low-abundance BRAF p.
Mutation prevalence increased progressively across Bethesda categories, ranging from 33% in Bethesda I to 89% in Bethesda VI.
This study developed a six-gene mutation detection panel composed of four multiplex droplet digital PCR (ddPCR) assays for thyroid nodule analysis.
Integration with cytology substantially reduces indeterminate diagnoses and may guide personalized management, including the avoidance of unnecessary surgery.