The gp63 gene locus, a target for genetic characterization of Leishmania belonging to subgenus Viannia.
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 (gp63-RFLP), and (ii) intra-genic PCR amplification coupled with restriction analysis (PCR-RFLP). 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 gp63-RFLP evidenced an extensive polymorphism and revealed specific markers for subgenus, species and geographical populations: congruence with MLEE was demonstrated statistically. The particular interest of gp63-RFLP 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 gp63-RFLP, but also allowed discrimination of the 2 subgenera (PCR alone) and all the species tested in the subgenus Viannia (PCR-RFLP). PCR-RFLP presents an important operational advantage as it allows genetic characterization of minute amounts of parasites, using Leishmania specific primers. The polymorphism revealed by gp63-RFLP and PCR-RFLP illustrates the very high genomic and genetic plasticity of gp63 genes.
Laboratory of Protozoology, Instituut voor Tropische Geneeskunde, Prins Leopold, Antwerpen, Belgium.
K
K
Victoir
A L
AL
Bañuls
J
J
Arevalo
A
A
Llanos-Cuentas
R
R
Hamers
S
S
Noël
S
S
De Doncker
D
D
Le Ray
M
M
Tibayrenc
J C
JC
Dujardin
D000818:Animals; D015139:Blotting, Southern; D004587:Electrophoresis, Agar Gel; D004588:Electrophoresis, Cellulose Acetate; D014644:Genetic Variation / Q000235:genetics; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics*; D007527:Isoenzymes / Q000032:analysis; D007891:Leishmania / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D007896:Leishmaniasis / Q000453:epidemiology* / Q000469:parasitology; D009693:Nucleic Acid Hybridization; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D015183:Restriction Mapping; D013020:South America / Q000453:epidemiology
D006023:Glycoproteins; D007527:Isoenzymes