Data/hora: 01/06/2024 01:03:38

Provedor de dados: 	Ciência Rural	
País: 	Brazil	
Título: 	Canine distemper virus detection by different methods of One-Step RT-qPCR	
Autores: 	Tozato,Claudia de Camargo; Zadra,Vívian Ferreira; Basso,Caroline Rodrigues; Araújo Junior,João Pessoa.	
Data: 	2016-09-01	
Ano: 	2016	
Palavras-chave: 	Canine distemper; Molecular diagnosis; One-Step RT-qPCR; Urine; Centrifugation.	
Resumo: 	ABSTRACT: Three commercial kits of One-Step RT-qPCR were evaluated for the molecular diagnosis of Canine Distemper Virus. Using the kit that showed better performance, two systems of Real-time RT-PCR (RT-qPCR) assays were tested and compared for analytical sensitivity to Canine Distemper Virus RNA detection: a One-Step RT-qPCR (system A) and a One-Step RT-qPCR combined with NESTED-qPCR (system B). Limits of detection for both systems were determined using a serial dilution of Canine Distemper Virus synthetic RNA or a positive urine sample. In addition, the same urine sample was tested using samples with prior centrifugation or ultracentrifugation. Commercial kits of One-Step RT-qPCR assays detected canine distemper virus RNA in 10 (100%) urine samples from symptomatic animals tested. The One-Step RT-qPCR kit that showed better results was used to evaluate the analytical sensitivity of the A and B systems. Limit of detection using synthetic RNA for the system A was 11 RNA copies µL-1 and 110 RNA copies µl-1 for first round System B. The second round of the NESTED-qPCR for System B had a limit of detection of 11 copies µl-1. Relationship between Ct values and RNA concentration was linear. The RNA extracted from the urine dilutions was detected in dilutions of 10-3 and10-2 by System A and B respectively. Urine centrifugation increased the analytical sensitivity of the test and proved to be useful for routine diagnostics. The One-Step RT-qPCR is a fast, sensitive and speciﬁc method for canine distemper routine diagnosis and research projects that require sensitive and quantitative methodology.	
Tipo: 	Info:eu-repo/semantics/article	
Idioma: 	Inglês	
Identificador: 	http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782016000901601	
Editor: 	Universidade Federal de Santa Maria	
Relação: 	10.1590/0103-8478cr20150932	
Formato: 	text/html	
Fonte: 	Ciência Rural  v.46 n.9 2016	
Fonte: 	info:eu-repo/semantics/openAccess