A novel DNA strip assay, GenoType MTBC, was evaluated for differentiation of Mycobacterium tuberculosis complex species from 77 positive liquid cultures in clinical practice. Species identification (M. tuberculosis [71 strains], Mycobacterium bovis subsp. bovis [5 strains], and Mycobacterium africanum subtype I [1 strain]) results were identical to conventional results. The sensitivity was slightly higher for this test than for the AccuProbe assay.