| Registro completo |
| Provedor de dados: |
Ciência Rural
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| País: |
Brazil
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| Título: |
Different methods of real-time PCR for detection of pseudorabies virus
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| 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
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| Data: |
2017-01-01
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| Ano: |
2017
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| Palavras-chave: |
Real time PCR
Pseudorabies
Diagnose
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| 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.
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| Tipo: |
Info:eu-repo/semantics/article
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| Idioma: |
Inglês
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| Identificador: |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782017000300452
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| Editor: |
Universidade Federal de Santa Maria
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| Relação: |
10.1590/0103-8478cr20160342
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| Formato: |
text/html
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| Fonte: |
Ciência Rural v.47 n.3 2017
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| Direitos: |
info:eu-repo/semantics/openAccess
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