Researchers at the University of Pittsburgh School of Medicine and the University of Pittsburgh Medical Center Hillman Cancer Center developed BiliSeq, a 28-gene DNA-based next-generation sequencing panel designed to improve diagnosis in bile duct strictures where imaging and pathology are often inconclusive. In a six-year prospective real-time study, BiliSeq detected about 82% of bile duct cancers compared with 44% using pathology alone. The team reported that combining BiliSeq with pathology increased cancer detection to nearly 90% while maintaining high specificity. The work tested upgraded BiliSeq versions (V2 and V3) on specimens spanning brushings, biopsies, and bile, totaling nearly 3,000 samples across more than 2,000 patients. Sensitivity and specificity improved further when the molecular assay was used alongside standard diagnostic workflows, with BiliSeqV2/V3 showing sensitivity of roughly 82% and specificity around 98% on its own, then rising to about 88% sensitivity when paired with pathology. The findings, published in Gastroenterology, underscore a move toward molecular confirmation to guide treatment decisions in this difficult cancer setting.