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An improved validated SYBR green-based real-time quantitative PCR assay for the detection of the Penaeus stylirostris densovirus in penaeid shrimp.
Encinas-García, Trinidad; Mendoza-Cano, Fernando; Enríquez-Espinoza, Tania; Luken-Vega, Leonardo; Vichido-Chávez, Rodrigo; Sánchez-Paz, Arturo.
Afiliação
  • Encinas-García T; Centro de Investigaciones Biológicas del Noroeste (CIBNOR), Laboratorio de Referencia, Análisis y Diagnóstico en Sanidad Acuícola, Hermosa 101, Col. Los Ángeles, Hermosillo, Sonora C.P. 83106, Mexico.
  • Mendoza-Cano F; Centro de Investigaciones Biológicas del Noroeste (CIBNOR), Laboratorio de Referencia, Análisis y Diagnóstico en Sanidad Acuícola, Hermosa 101, Col. Los Ángeles, Hermosillo, Sonora C.P. 83106, Mexico.
  • Enríquez-Espinoza T; Centro de Investigaciones Biológicas del Noroeste (CIBNOR), Laboratorio de Referencia, Análisis y Diagnóstico en Sanidad Acuícola, Hermosa 101, Col. Los Ángeles, Hermosillo, Sonora C.P. 83106, Mexico.
  • Luken-Vega L; Departamento de Investigaciones Científicas y Tecnológicas, Universidad de Sonora, Av. Colosio s/n, entre Sahuaripa y Reforma, Hermosillo, Sonora 83000, Mexico.
  • Vichido-Chávez R; Departamento de Diagnóstico Acuícola y Pesquero, Centro Nacional de Servicios de Constatación en Salud Animal (CENAPA), Carr. Fed. Cuernavaca Cuautla 8534 Colonia Progreso, Jiutepec, Morelos C.P. 62550, Mexico.
  • Sánchez-Paz A; Centro de Investigaciones Biológicas del Noroeste (CIBNOR), Laboratorio de Referencia, Análisis y Diagnóstico en Sanidad Acuícola, Hermosa 101, Col. Los Ángeles, Hermosillo, Sonora C.P. 83106, Mexico. Electronic address: asanchez04@cibnor.mx.
J Virol Methods ; 212: 53-8, 2015 Feb.
Article em En | MEDLINE | ID: mdl-25445798
The Penaeus stylirostris densovirus (PstDV) (also known as infectious hypodermal and hematopoietic necrosis virus, IHHNV), one of the major shrimp pathogens, has a worldwide distribution in farmed and wild shrimp populations. Outbreaks of IHHNV have been associated with substantial economic losses which are accompanied by a negative social impact. Current diagnostic PCR tests may result in false-positive results as several parts of PstDV genome may be endogenized in the nuclear genome of the shrimp P. stylirostris. A one-step qPCR SYBR-Green based quantitative real-time polymerase chain reaction (qPCR) assay to detect different isolates of the IHHNV in shrimp samples was developed. The detection limit of the assay was 81 viral copies of targeted DNA per reaction. The specificity of the assay was evaluated by melting curve analysis, which showed that the IHHNV product generated a single melt peak at 81.4±0.044°C. The assay was more sensitive than conventional PCR. The standardized PCR was shown to be highly sensible, specific, robust, and reproducible, which makes it an economical and powerful tool for both diagnostic applications and general research of IHHNV.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Virologia / Densovirus / Penaeidae / Reação em Cadeia da Polimerase em Tempo Real Tipo de estudo: Diagnostic_studies / Evaluation_studies Limite: Animals Idioma: En Revista: J Virol Methods Ano de publicação: 2015 Tipo de documento: Article País de afiliação: México País de publicação: Holanda
Texto completo
Adicionar na Minha BVS
Imprimir
XML
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Virologia / Densovirus / Penaeidae / Reação em Cadeia da Polimerase em Tempo Real Tipo de estudo: Diagnostic_studies / Evaluation_studies Limite: Animals Idioma: En Revista: J Virol Methods Ano de publicação: 2015 Tipo de documento: Article País de afiliação: México País de publicação: Holanda