Visceral leishmaniasis : a practical strategy for quantitative molecular diagnosis in naturally infected dogs.

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dc.contributor.authorCaldas, Sérgio
dc.contributor.authorMarcelino, Andreza Pain
dc.contributor.authorFaria, Gilson
dc.contributor.authorSilva, Fernanda de Oliveira
dc.contributor.authorAtaide, Ana Caroline Zampiroli
dc.contributor.authorCunha, Lucas Maciel
dc.contributor.authorBahia, Maria Terezinha
dc.contributor.authorPaz, Gustavo Fontes
dc.contributor.authorGontijo, Célia Maria Ferreira
dc.date.accessioned2021-11-22T15:10:39Z
dc.date.available2021-11-22T15:10:39Z
dc.date.issued2020pt_BR
dc.description.abstractThe diagnosis of canine visceral leishmaniasis (CVL) has been a problem for public health services due to the variety of clinical signs similar to other diseases and low sensitivity and specificity of available tests. In this sense, our main objective was to develop a simple, rapid, and accurate quantitative real-time PCR (qPCR) diagnosis for CVL. Thus, low-invasive samples from bone marrow (BM), popliteal lymph nodes (PLN), and conjunctival swabs (CS) were selected from negative and VL-positive dogs, using as gold standard, immunological and parasitological tests performed with different tissues. Oligonucleotides for Leishmania infantum kDNA were designed and the limit of quantification and amplification efficiency of the qPCR were determined using tissue-specific standards produced with DNA from those different tissues, mixed with DNA from a known amount of L. infantum promastigotes. Endogenous control was used to validate a comparative Ct method, and tissue parasite concentrations were estimated by comparison with tissue-specific reference standard samples. The overall analysis of the qPCR data suggests the following ranking for tissue choice: PLN > BM > CS. Finally, we have concluded that this molecular approach simplifies and accelerates the quantitative diagnostic process because it is easy to perform, requiring no DNA dosing or standard curve application, and it shows good diagnostic parameters, especially when using popliteal lymph node samples.pt_BR
dc.identifier.citationCALDAS, S. et al. Visceral leishmaniasis: a practical strategy for quantitative molecular diagnosis in naturally infected dogs. Parasitology Research, v. 119, p. 1683-1690, 2020. Disponível em: <https://link.springer.com/article/10.1007%2Fs00436-020-06654-y>. Acesso em: 10 jun. 2021.pt_BR
dc.identifier.doihttps://doi.org/10.1007/s00436-020-06654-ypt_BR
dc.identifier.issn1432-1955
dc.identifier.urihttp://www.repositorio.ufop.br/jspui/handle/123456789/14000
dc.identifier.uri2https://link.springer.com/article/10.1007%2Fs00436-020-06654-ypt_BR
dc.language.isoen_USpt_BR
dc.rightsrestritopt_BR
dc.subjectParasitespt_BR
dc.subjectQuantitative real time PCRpt_BR
dc.titleVisceral leishmaniasis : a practical strategy for quantitative molecular diagnosis in naturally infected dogs.pt_BR
dc.typeArtigo publicado em periodicopt_BR
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