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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Detection of Leishmania infantum DNA in conjunctival swabs of cats by quantitative real-time PCR

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Author(s):
Benassi, Julia Cristina ; Benvenga, Graziella U. ; Ferreira, Helena Lage ; Pereira, Vanessa F. ; Keid, Lara B. ; Soares, Rodrigo ; Ferreira de Sousa Oliveira, Tricia Maria
Total Authors: 7
Document type: Journal article
Source: Experimental Parasitology; v. 177, p. 93-97, JUN 2017.
Web of Science Citations: 9
Abstract

Although some studies have investigated the potential role of cats as a reservoir for Leishmania, their role in the epidemiology of visceral leishmaniasis (VL) is still poorly understood. Molecular diagnostic techniques are an important tool in VL diagnosis, and PCR shows high sensitivity and specificity for Leishmania spp. detection. Quantitative real-time PCR (qPCR) is a method that permits quantitative analysis of a large number of samples, resulting in more sensitive, accurate, and reproducible measurements of specific DNA present in the sample. This study compared real-time PCR (qPCR) and conventional PCR (cPCR) for detection of Leishmania spp. in blood and conjunctival swab (CS) samples of healthy cats from a non-endemic area in the state of Sao Paulo, Brazil. Of all CS samples, 1.85% (2/108) were positive for Leishmania spp. by both cPCR as qPCR (kappa index = 1), indicating excellent agreement between the two methods. The DNA from the two CS-cPCR- and CS-qPCR-positive samples was further tested with a PCR test amplifying the Leishmania spp. discriminative rRNA internal transcribed spacer 1 (ITS 1), of which one sample generated a 300-350-bp DNA fragment whose size varies according to the Leishmania species. Following sequencing, the fragment showed 100% similarity to a GenBank L. infantum sequence obtained from a cat in Italy. In conclusion, the association of qPCR and CS proved to be effective for detection of Leishmania in cats. Conjunctival swab samples were shown to be a practical and better alternative to blood samples and may be useful in the diagnosis and studies of feline leishmaniasis. (C) 2017 Elsevier Inc. All rights reserved. (AU)

FAPESP's process: 11/00147-6 - Conjunctival swab samples for canine leishmaniasis detection by PCR
Grantee:Trícia Maria Ferreira de Sousa Oliveira
Support Opportunities: Regular Research Grants