PCR as a rapid and sensitive tool in the diagnosis of human and canine leishmaniasis using Leishmania donovani s.l.-specific kinetoplastid primers.
This study was performed in order to test the efficacy of a new polymerase chain reaction (PCR) assay for the diagnosis of both human and canine leishmaniasis caused by Leishmania infantum. The new primers were developed on the basis of a complete DNA sequence of the L. infantum kinetoplast minicircle. Specificity and sensitivity were evaluated by testing bone marrow spots on filter paper and skin biopsy samples, and the PCR results were compared to data from in vitro cultures. Leishmania strains from different foci, as well as other trypanosomatids and opportunistic pathogenic micro-organisms, were also included in this study. The results show that the primers are highly specific, detecting only L. donovani s.l. DNA, and sensitive for the detection of parasite DNA in biological samples from three different geographical regions of Portugal (north, centre and south) and from Brazil.
Cortes
S
S
Unidade de Leishmanioses, Centro Malária Outras Doenças Tropicais, Instituto de Higiene e Medicina Tropical (IHMT), Universidade Nova de Lisboa, Rua da Junqueira 96, 1349-008 Lisboa, Portugal.
Rolão
N
N
Ramada
J
J
Campino
L
L
D000818:Animals; D017931:DNA Primers; D018105:DNA, Kinetoplast; D004283:Dog Diseases / Q000175:diagnosis* / Q000469:parasitology; D004285:Dogs; D006801:Humans; D018314:Leishmania infantum / Q000235:genetics*; D007898:Leishmaniasis, Visceral / Q000175:diagnosis* / Q000662:veterinary*; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards / Q000662:veterinary; D012680:Sensitivity and Specificity
D017931:DNA Primers; D018105:DNA, Kinetoplast