Forty-six independent isolates of Leishmania subgenus were examined by restriction fragment length polymorphism (RFLP) analyses of polymerase chain reaction (PCR)-amplified mimexon sequences. The results were consistent with grouping of these isolates into species complexes based on sequence polymorphism of single-copy genes. Mimexon RFLP profiles specific to Leishmania enriettii, Leishmania tarentolae, and Leishmania gerbilli were obtained for the first time. The mimexon PCR-RFLP method is thus further confirmed as a simple, sensitive, and specific method useful for discriminating Leishmania isolates. (c) 2007 Elsevier Inc. All rights reserved.