Author | Caldas, Sérgio | |
Author | Marcelino, Andreza P. | |
Author | Faria, Gilson | |
Author | Silva, Fernanda de Oliveira | |
Author | Ataide, Ana Caroline Zampiroli | |
Author | Cunha, Lucas Maciel | |
Author | Bahia, Maria Terezinha | |
Author | Paz, Gustavo Fontes | |
Author | Gontijo, Célia M. F. | |
Access date | 2020-12-15T19:09:20Z | |
Available date | 2020-12-15T19:09:20Z | |
Document date | 2020 | |
Citation | CALDAS, Sérgio et al. Visceral leishmaniasis: a practical strategy for quantitative molecular diagnosis in naturally infected dogs. Parasitology Research, v. 119, n. 5, p. 1683-1690, 2020. | pt_BR |
ISSN | 0932-0113 | pt_BR |
URI | https://www.arca.fiocruz.br/handle/icict/44917 | |
Language | eng | pt_BR |
Publisher | Springer | pt_BR |
Rights | restricted access | pt_BR |
Title | Visceral leishmaniasis: a practical strategy for quantitative molecular diagnosis in naturally infected dogs | pt_BR |
Type | Article | pt_BR |
DOI | 10.1007/s00436-020-06654-y | |
Abstract | The 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 |
Affilliation | Fundação Ezequiel Dias. Diretoria de Pesquisa e Desenvolvimento. Serviço de Biotecnologia e Saúde. Belo Horizonte, MG, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil. | pt_BR |
Affilliation | Fundação Ezequiel Dias. Diretoria de Pesquisa e Desenvolvimento. Serviço de Biotecnologia e Saúde. Belo Horizonte, MG, Brasil. | pt_BR |
Affilliation | Fundação Ezequiel Dias. Diretoria de Pesquisa e Desenvolvimento. Serviço de Biotecnologia e Saúde. Belo Horizonte, MG, Brasil. | pt_BR |
Affilliation | Fundação Ezequiel Dias. Diretoria de Pesquisa e Desenvolvimento. Serviço de Biotecnologia e Saúde. Belo Horizonte, MG, Brasil. | pt_BR |
Affilliation | Fundação Ezequiel Dias. Diretoria de Pesquisa e Desenvolvimento. Serviço de Biotecnologia e Saúde. Belo Horizonte, MG, Brasil. | pt_BR |
Affilliation | Universidade Federal de Ouro Preto. Escola de Medicina & Núcleo de Pesquisas em Ciências Biológicas. Laboratório de Doenças Parasitárias. Ouro Preto, MG, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto René Rachou. Belo Horizonte, MG, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto René Rachou. Belo Horizonte, MG, Brasil. | pt_BR |
Subject | Molecular diagnosis | pt_BR |
Subject | Parasites | pt_BR |
Subject | Quantitative real-time PCR | pt_BR |
Subject | Visceral leishmaniasis | pt_BR |