The gp63 gene locus, a target for genetic characterization of **Leishmania** belonging to subgenus **Viannia**
Faculty of Pharmaceutical, Biomedical and Veterinary Sciences . Biomedical Sciences
Parasitology. - London, 1908, currens
, p. 1-13
In the present study the gp63 gene locus was used as a target for genetic characterization of Leishmania parasites by 2 methods: (i) RFLP analysis with several restriction enzymes (gp63RFLP), and (ii) intra-genic PCR amplification coupled with restriction analysis (PCRRFLP). Both methods were applied to a large number of natural isolates belonging to 4 species of the subgenus Viannia, namely L. (V.) braziliensis, L. (V.) peruviana, L. (V.) guyanensis and L. (V.) lainsoni: reference stocks of subgenus Leishmania were included as outgroups. Multilocus isoenzyme typing (MLEE) was used as a reference. On the one hand gp63RFLP evidenced an extensive polymorphism and revealed specific markers for subgenus, species and geographical populations: congruence with MLEE was demonstrated statistically. The particular interest of gp63RFLP was illustrated by infra-specific polymorphism, because of the possible relationship with phenotype diversity. On the other hand intra-genic amplification was less resolutive than gp63RFLP, but also allowed discrimination of the 2 subgenera (PCR alone) and all the species tested in the subgenus Viannia (PCRRFLP). PCRRFLP presents an important operational advantage as it allows genetic characterization of minute amounts of parasites, using Leishmania specific primers. The polymorphism revealed by gp63RFLP and PCRRFLP illustrates the very high genomic and genetic plasticity of gp63 genes.