Leishmania (Viannia) subgenus kDNA amplification for the diagnosis of mucosal leishmaniasis.
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.
Disch
Jolande
J
Laboratory of Clinical Research, Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz (FIOCRUZ), Belo Horizonte, MG 30190-002, Brazil.
Pedras
Mariana Junqueira
MJ
Orsini
Marcela
M
Pirmez
Claude
C
de Oliveira
Maria Cláudia
MC
Castro
Marcelo
M
Rabello
Ana
A
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
D018105:DNA, Kinetoplast