The mango fruit fly, Ceratitis cosyra, is a major agricultural pest affecting mango production in sub-Saharan Africa. Morphological differences between C. cosyra and four closely related Ceratitis (Ceratalaspis) species (C. discussa, C. quinaria, C. silvestrii, C. striatella) are subtle, so that reliable separation often requires specialized expertise. Additionally, a previous study showed cryptic genetic variation in C. cosyra with microsatellite genotypic clusters also occurring in sympatry. DNA barcoding further supported cryptic genetic variation in C. cosyra due to outlier DNA sequences. This study aimed to verify whether DNA barcoding can profitably be used to (a) resolve the two main microsatellite genotypic clusters of C. cosyra, and (b) separate C. cosyra from C. discussa, C. quinaria, C. silvestrii, and C. striatella. A subset of 48 C. cosyra specimens previously assigned to the two microsatellite genotypic clusters was subjected to DNA barcoding. This dataset was integrated with 83 public DNA barcodes of C. cosyra, C. discussa, C. quinaria, C. silvestrii, and C. striatella from the Barcode of Life Data Systems (BOLD). Neighbour joining tree (K2P distance) separated the two C. cosyra genotypic clusters and also resolved properly supported groups corresponding to (a) C. quinaria and C. silvestrii, (b) C. discussa (only 2 specimens considered) and possibly (c) C. striatella (all specimens but one included in the same clade). These results further support the hypothesis of cryptic speciation in the mango fruit fly and suggest that DNA barcoding represents a suitable tool for the identification of C. cosyra and of the closely related Ceratitis (Ceratalaspis) species. (Texte intégral)