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Application and development of a TaqMan-based real-time PCR assay for rapid detection of snakehead vesiculovirus.
Gong, Cuiping; Zhu, Panpan; Ye, Jiaxin; Lou, Jianfeng; Zhang, Liwen; Liu, Xiaodan; Kong, Weiguang.
Afiliación
  • Gong C; Huzhou Academy of Agricultural Sciences, Huzhou Municipal Bureau of Agriculture and Rural Affairs, Huzhou 313000, China.
  • Zhu P; College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
  • Ye J; College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
  • Lou J; Huzhou Academy of Agricultural Sciences, Huzhou Municipal Bureau of Agriculture and Rural Affairs, Huzhou 313000, China.
  • Zhang L; College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
  • Liu X; College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
  • Kong W; International Research Laboratory of Prevention and Control of Important Animal Infectious Diseases and Zoonotic Diseases of Jiangsu Higher Education Institutions, Yangzhou University, Yangzhou 225009, China.
FEMS Microbiol Lett ; 3712024 Jan 09.
Article en En | MEDLINE | ID: mdl-38460951
ABSTRACT
Snakehead vesiculovirus (SHVV) is one of the primary pathogens responsible for viral diseases in the snakehead fish. A TaqMan-based real-time PCR assay was established for the rapid detection and quantification of SHVV in this study. Specific primers and fluorescent probes were designed for phosphoprotein (P) gene, and after optimizing the reaction conditions, the results indicated that the detection limit of this method could reach 37.1 copies, representing a 100-fold increase in detection sensitivity compared to RT-PCR. The specificity testing results revealed that this method exhibited no cross-reactivity with ISKNV, LMBV, RSIV, RGNNV, GCRV, and CyHV-2. Repetition experiments demonstrated that both intra-batch and inter-batch coefficients of variation were not higher than 1.66%. Through in vitro infection experiments monitoring the quantitative changes of SHVV in different tissues, the results indicated that the liver and spleen exhibited the highest viral load at 3 poi. The TaqMan-based real-time PCR method established in this study exhibits high sensitivity, excellent specificity, and strong reproducibility. It can be employed for rapid detection and viral load monitoring of SHVV, thus providing a robust tool for the clinical diagnosis and pathogen research of SHVV.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Perciformes / Infecciones por Rhabdoviridae / Enfermedades de los Peces / Iridoviridae Límite: Animals Idioma: En Revista: FEMS Microbiol Lett Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Perciformes / Infecciones por Rhabdoviridae / Enfermedades de los Peces / Iridoviridae Límite: Animals Idioma: En Revista: FEMS Microbiol Lett Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Reino Unido