Turcinoemacheilus ekmekciae, new species, from upper Euphrates and Tigris drainages is distinguished from other species of Turcinoemacheilus in Western Asia by having a dark stripe broader than the eye diameter along the lateral line, rarely possessing roundish blotches, 5–6 mandibular pores in mandibular canal, a comperatvely smaller head, a deeper body, and a greater pre-pelvic distance. Our specimens collected from the upper Great Zab, near the type locality of Turcinoemacheilus kosswigi, showed notable genetic divergence (a minimum K2P of 3.3%) from sequences reported as T. kosswigi in previous studies. Despite morphological similarities, this molecular difference suggests that the populations analysed in previous studies may represent a potential new species of Turcinoemacheilus, which we tentatively named as Turcinoemacheius cf. kosswigi. Molecular data also suggest that T. ekmekciae is characterized by a minimum K2P distance of 3.5% from Turcinoemacheilus minimus and T. cf. kosswigi. The three methods for species delimitation (assemble species by automatic partitioning [ASAP], Poisson tree processes [PTP], and multi-rate PTP [mPTP]) that were utilized for testing species assignments consistently identified our test group as a distinct species.