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Evaluation of urine for Leishmania infantum DNA detection by real-time quantitative PCR.
Pessoa-E-Silva, Rômulo; Mendonça Trajano-Silva, Lays Adrianne; Lopes da Silva, Maria Almerice; da Cunha Gonçalves-de-Albuquerque, Suênia; de Goes, Tayná Correia; Silva de Morais, Rayana Carla; Lopes de Melo, Fábio; de Paiva-Cavalcanti, Milena.
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  • Pessoa-E-Silva R; Aggeu Magalhães Research Center, Av. Moraes Rego, Cidade Universitária, CEP 50670-420 Recife, PE, Brazil; Public Health's Central Laboratory Dr. Milton Bezerra Sobral (LACEN-PE), Praça Oswaldo Cruz, S/N, Soledade, CEP 50050-215 Recife, PE, Brazil. Electronic address: romulops12@gmail.com.
  • Mendonça Trajano-Silva LA; Aggeu Magalhães Research Center, Av. Moraes Rego, Cidade Universitária, CEP 50670-420 Recife, PE, Brazil. Electronic address: laystrajano@gmail.com.
  • Lopes da Silva MA; Aggeu Magalhães Research Center, Av. Moraes Rego, Cidade Universitária, CEP 50670-420 Recife, PE, Brazil. Electronic address: almerice@cpqam.fiocruz.br.
  • da Cunha Gonçalves-de-Albuquerque S; Aggeu Magalhães Research Center, Av. Moraes Rego, Cidade Universitária, CEP 50670-420 Recife, PE, Brazil; Public Health's Central Laboratory Dr. Milton Bezerra Sobral (LACEN-PE), Praça Oswaldo Cruz, S/N, Soledade, CEP 50050-215 Recife, PE, Brazil. Electronic address: sueniaaa@hotmail.com.
  • de Goes TC; Aggeu Magalhães Research Center, Av. Moraes Rego, Cidade Universitária, CEP 50670-420 Recife, PE, Brazil. Electronic address: taynacgoes@hotmail.com.
  • Silva de Morais RC; Aggeu Magalhães Research Center, Av. Moraes Rego, Cidade Universitária, CEP 50670-420 Recife, PE, Brazil. Electronic address: rayanacarla_m@hotmail.com.
  • Lopes de Melo F; Aggeu Magalhães Research Center, Av. Moraes Rego, Cidade Universitária, CEP 50670-420 Recife, PE, Brazil. Electronic address: fabio@cpqam.fiocruz.br.
  • de Paiva-Cavalcanti M; Aggeu Magalhães Research Center, Av. Moraes Rego, Cidade Universitária, CEP 50670-420 Recife, PE, Brazil. Electronic address: mp@cpqam.fiocruz.br.
J Microbiol Methods ; 131: 34-41, 2016 12.
Article en En | MEDLINE | ID: mdl-27713020
The availability of some sorts of biological samples which require noninvasive collection methods has led to an even greater interest in applying molecular biology on visceral leishmaniasis (VL) diagnosis, since these samples increase the safety and comfort of both patients and health professionals. In this context, this work aimed to evaluate the suitability of the urine as a specimen for Leishmania infantum kinetoplast DNA detection by real-time quantitative PCR (qPCR). Subsequent to the reproducibility analysis, the detection limit of the qPCR assay was set at 5fg (~0.025 parasites) per µL of urine. From the comparative analysis performed with a set of diagnostic criteria (serological and molecular reference tests), concordance value of 96.08% was obtained (VL-suspected and HIV/AIDS patients, n=51) (P>0.05). Kappa coefficient (95% CI) indicated a good agreement between the test and the set of diagnostic criteria (k=0.778±0.151). The detection of Leishmania DNA in urine by qPCR was possible in untreated individuals, and in those with or without suggestive renal impairment. Fast depletion of the parasite's DNA in urine after treatment (from one dose of meglumine antimoniate) was suggested by negative qPCR results, thus indicating it as a potential alternative specimen to follow up the efficacy of therapeutic approaches. Even when evaluated in a clinically heterogeneous set of patients, the urine showed good prospect as sample for VL diagnosis by qPCR, also indicating a good negative predictive value for untreated suspected patients.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Orina / Leishmania infantum / ADN de Cinetoplasto / Técnicas de Diagnóstico Molecular / Reacción en Cadena en Tiempo Real de la Polimerasa / Leishmaniasis Visceral Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Adolescent / Adult / Aged / Child / Female / Humans / Male / Middle aged País/Región como asunto: America do sul / Brasil Idioma: En Revista: J Microbiol Methods Año: 2016 Tipo del documento: Article Pais de publicación: Países Bajos
Texto completo
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Imprimir
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Orina / Leishmania infantum / ADN de Cinetoplasto / Técnicas de Diagnóstico Molecular / Reacción en Cadena en Tiempo Real de la Polimerasa / Leishmaniasis Visceral Tipo de estudio: Diagnostic_studies / Prognostic_studies Límite: Adolescent / Adult / Aged / Child / Female / Humans / Male / Middle aged País/Región como asunto: America do sul / Brasil Idioma: En Revista: J Microbiol Methods Año: 2016 Tipo del documento: Article Pais de publicación: Países Bajos