Journal
VETERINARY PARASITOLOGY
Volume 197, Issue 3-4, Pages 498-503Publisher
ELSEVIER
DOI: 10.1016/j.vetpar.2013.07.006
Keywords
Leishmania infantum; Molecular diagnostic; PCR; qPCR; snPCR; Skin; Spleen; Canine visceral leishmaniasis
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Funding
- DECIT/MS/CNPq/BR [576062/2008-1]
- FAPEMIG/BR [CBB - APQ-3073-4.01/07]
- CNPq/BR [472554/2007-7]
- PPSUS/MS/CNPq/FAPEMIG/SES-MG [CBB-APQ-00356-10]
- Federal University of Ouro Preto
- Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)
- CAPES/PNPD
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Polymerase chain reaction (PCR) and its variations represent highly sensitive and specific methods for Leishmania DNA detection and subsequent canine visceral leishmaniasis (CVL) diagnosis. The aim of this work was to compare three different molecular diagnosis techniques (conventional PCR [cPCR], seminested PCR [snPCR], and quantitative PCR [qPCR]) in samples of skin and spleen from 60 seropositive dogs by immunofluorescence antibody test and enzyme-linked immunosorbent assay. Parasitological analysis was conducted by culture of bone marrow aspirate and optical microscopic assessment of ear skin and spleen samples stained with Giemsa, the standard tests for CVL diagnosis. The primers L150/L152 and LINR4/LIN17/LIN19 were used to amplify the conserved region of the Leishmania kDNA minicircle in the cPCR, and snPCR and qPCR were performed using the DNA polymerase gene (DNA pol alpha) primers from Leishmania infantum. The parasitological analysis revealed parasites in 61.7% of the samples. Sensitivities were 89.2%, 86.5%, and 973% in the skin and 81.1%, 94.6%, and 100.0% in spleen samples used for cPCR, snPCR, and qPCR, respectively. We demonstrated that the qPCR method was the best technique to detect L infantum in both skin and spleen samples. However, we recommend the use of skin due to the high sensitivity and sampling being less invasive. (C) 2013 Elsevier B.V. All rights reserved.
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