A new species of Nannocharax is described from the Luongo and Kalungwishi Rivers, tributaries of the Luapula River in the northeastern region of Zambia. The new species differs from its congeners by a combination of characters, such as the body coloration pattern formed by a series of one-scale-wide vertical bars, a small caudal-peduncle spot surrounded by a light clear area at the base of the middle caudal-fin rays, and a low number of scales in the circumpeduncular series and lateral line series. The new species is also distinguished from other members of the Nannocharax multifasciatus species-group by genetic distances ranging from 10.3% to 11.6% with DNA barcoding. Likewise, distance and coalescent molecular species delimitation approaches recovered the new species as an independent operational taxonomic unit. Newly generated hypotheses of phylogenetic relationships based on maximum likelihood and Bayesian inference for the new taxon are provided.