title: Leishmania (Viannia) subgenus kDNA amplification for the diagnosis of mucosal leishmaniasis.
abstract: The utility of 2 polymerase chain reaction (PCR)-based assays amplifying genus or Viannia subgenus Leishmania minicircle kDNA for the diagnostics of ML was assessed. The Viannia subgenus product was yielded after PCR from isolates of L. (Viannia) braziliensis, L. (Viannia) colombiensis, and L. (Viannia) guyanensis, whereas no product was obtained with the non-Viannia-pertaining species: L. (Leishmania) amazonensis, L. (Leishmania) donovani, and L. (Leishmania) chagasi. With both assays, 11 of 13 (86.4%) patients with confirmed ML could be identified, whereas only 2 (16.7%) of these patients were positive by microscopy. All amplified genus-specific products gave a positive signal by hybridization with a Leishmania (Viannia) subgenus-specific radioactive probe. The Viannia subgenus-specific kDNA PCR represents a sensitive and specific tool for the diagnosis of ML, remarkably improving the sensitivity of parasitological methods and offering an alternative for the radioactive-dependent assays for subgenus characterization.
authors.affiliation: Laboratory of Clinical Research, Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz (FIOCRUZ), Belo Horizonte, MG 30190-002, Brazil.
authors.forename: Jolande
authors.initials: J
authors.lastname: Disch
authors.forename: Mariana Junqueira
authors.initials: MJ
authors.lastname: Pedras
authors.forename: Marcela
authors.initials: M
authors.lastname: Orsini
authors.forename: Claude
authors.initials: C
authors.lastname: Pirmez
authors.forename: Maria Cláudia
authors.initials: MC
authors.lastname: de Oliveira
authors.forename: Marcelo
authors.initials: M
authors.lastname: Castro
authors.forename: Ana
authors.initials: A
authors.lastname: Rabello
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D018105:DNA, Kinetoplast / Q000235:genetics*; D005260:Female; D006801:Humans; D007891:Leishmania / Q000145:classification / Q000235:genetics*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis*; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D018105:DNA, Kinetoplast