title: Direct identification of Leishmania species in biopsies from patients with American tegumentary leishmaniasis.
abstract: Accurate identification of Leishmania species is important for monitoring clinical outcome, adequately targeting treatment, and evaluation of epidemiological risk in tegumentary leishmaniasis. This is especially the case in regions where several species coexist and for travel medicine where the geographical source of infection is not always obvious. Species identification presently depends on parasite isolation, which is not very sensitive and not necessarily representative of parasites actually present in human tissues. We evaluated a polymerase chain reaction (PCR) assay combining amplification of the gp63 genes and restriction fragment length polymorphism (RFLP) analysis (gp63 PCR-RFLP) for direct Leishmania species-identification in tissues collected from Peruvian patients in 1999. By comparison with a kinetoplast DNA-based PCR, our PCR assay showed a detection sensitivity of 85%. Three species were encountered among patient samples, Leishmania (Viannia) braziliensis, L. (V.) peruviana and L. (V.) guyanensis, and their frequency and geographical distribution corresponded to earlier epidemiological studies of leishmaniasis in Peru. However, unexpected results raised questions about (i) the contribution of human migration to the emergence of new foci of given species, (ii) the pathogenicity of some species, and (iii) the frequency of mixed or hybrid infections.
authors.affiliation: Prins Leopold Instituut voor Tropische Geneeskunde, Nationalestraat 155, B-2000 Antwerpen, Belgium.
authors.forename: Kathleen
authors.initials: K
authors.lastname: Victoir
authors.forename: Simonne
authors.initials: S
authors.lastname: De Doncker
authors.forename: Leyda
authors.initials: L
authors.lastname: Cabrera
authors.forename: Eugenia
authors.initials: E
authors.lastname: Alvarez
authors.forename: Jorge
authors.initials: J
authors.lastname: Arevalo
authors.forename: Alejandro
authors.initials: A
authors.lastname: Llanos-Cuentas
authors.forename: Dominique
authors.initials: D
authors.lastname: Le Ray
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Dujardin
mesh_terms: D000818:Animals; D015342:DNA Probes; D016054:DNA, Protozoan; D006801:Humans; D007892:Leishmania braziliensis / Q000145:classification / Q000302:isolation & purification*; D007896:Leishmaniasis / Q000175:diagnosis* / Q000469:parasitology; D010274:Parasitology / Q000379:methods / Q000592:standards; D010568:Peru; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity
chemical_list: D015342:DNA Probes; D016054:DNA, Protozoan