The performance of the Capilia TB-Neo assay, a new-generation assay, was assessed by determining its sensitivity, specificity, reproducibility, and cross-reaction with contaminating organisms. The sensitivity and specificity were 99.2 and 96.4% and 89.3 and 100% in pure and mixed-culture isolates, respectively. The kappa statistic was 95.0 and 77.9% in pure and mixed culture isolates, respectively. There was no cross-reaction with contaminating organisms.