Data/hora: 06/06/2024 06:26:40

Provedor de dados: | 65
País: | Brazil
Título: | Different methods of real-time PCR for detection of pseudorabies virus
Autores: | Nonaka,Carolina Kymie Vasques; Fonseca Junior,Antônio Augusto; Guedes,Estefânia Oliveira; D´Ambros,Régia Maria; Lima,Graciela Kunrath; Camargos,Marcelo Fernandes; Heinemann,Marcos Bryan.
Data: | 2017-01-01
Ano: | 2017
Palavras-chave: | Real time PCR; Pseudorabies; Diagnose.
Resumo: | ABSTRACT: Pseudorabies (PR) is a highly contagious viral disease of great animal health and economic importance in swine industry. The aim of this study was to evaluate different genomic regions, real-time PCR chemistries and equipment for the molecular diagnosis of PR. Eight primer pairs targeting four genes (gB, gC, gE, gD), three different qPCR chemistries (SybrGreen, hydrolysis probes and plexor) and two equipment (ABI7500, Rotorgene 3000) were evaluated. Oligonucleotides targeting gB using hydrolysis probes showed the best performance after evaluating efficiency (99%), the detection limit (10-1.5 TCID50 mL-1) and diagnostic sensitivity and; therefore, those primers were selected for performance verification factors such as repeatability, reproducibility and robustness (1.39% variance between days, 24% variance between analysts and 4.07% variance in analysis error). The qPCR standardized and validated in this research proved to be reliable for the diagnosis of PR and may be used in diagnostic laboratories that follow ISO 17025 and ISO 16140.
Tipo: | Info:eu-repo/semantics/article
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782017000300452
Editor: | Universidade Federal de Santa Maria
Relação: | 10.1590/0103-8478cr20160342
Formato: | text/html
Fonte: | Ciência Rural  v.47 n.3 2017
Fonte: | info:eu-repo/semantics/openAccess