Differentiation of Leishmania (Viannia) panamensis and Leishmania (V.) guyanensis using BccI for hsp70 PCR-RFLP.
Leishmania panamensis and Leishmania guyanensis are two species of the subgenus Viannia that are genetically very similar. Both parasites are usually associated with cutaneous leishmaniasis, but also have the potential to cause the mucocutaneous form of the disease. In addition, the study of foci and consequently the identification of vectors and probable reservoirs involved in transmission require a correct differentiation between both species, which is important at epidemiological level. We explored the possibility of identifying these species by using restriction fragment length polymorphisms (RFLP) in the gene coding for heat-shock protein 70 (hsp70). Previously, an hsp70 PCR-RFLP assay proved to be very effective in differentiating other Leishmania species when HaeIII is used as restriction enzyme. Based on hsp70 sequences analysis, BccI was found to generate species-specific fragments that can easily be recognized by agarose gel electrophoresis. Using the analysis of biopsies, scrapings, and parasite isolates previously grouped in a cluster comprising both L. panamensis and L. guyanensis, we showed that our approach allowed differentiation of both entities. This offers the possibility not only for identification of parasites in biological samples, but also to apply molecular epidemiology in certain countries of the New World, where several Leishmania species could coexist.
Instituto de Medicina Tropical Pedro Kourí, Apartado Postal 601, Marianao 13, La Habana, Cuba.
Ana Margarita
AM
Montalvo Alvarez
Jorge Fraga
JF
Nodarse
Ivón Montano
IM
Goodridge
Lianet Monzote
LM
Fidalgo
Marcel
M
Marin
Gert
G
Van Der Auwera
Jean-Claude
JC
Dujardin
Iván Darío Velez
ID
Bernal
Carlos
C
Muskus
D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D006801:Humans; D018334:Leishmania guyanensis / Q000145:classification / Q000235:genetics*; D016773:Leishmaniasis, Cutaneous / Q000235:genetics / Q000469:parasitology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA; D013045:Species Specificity
D018840:HSP70 Heat-Shock Proteins