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Validation of specific quantitative real-time RT-PCR assay panel for Infectious Bronchitis using synthetic DNA standards and clinical specimens.
Mo, Jongseo; Angelichio, Michael; Gow, Lisa; Leathers, Valerie; Jackwood, Mark W.
Afiliación
  • Mo J; Poultry Diagnostic and Research Center, Department of Population Health, College of Veterinary Medicine, University of Georgia, Athens GA, United States.
  • Angelichio M; IDEXX Laboratories, Inc., One IDEXX Drive, Westbrook, ME, United States.
  • Gow L; IDEXX Laboratories, Inc., One IDEXX Drive, Westbrook, ME, United States.
  • Leathers V; IDEXX Laboratories, Inc., One IDEXX Drive, Westbrook, ME, United States.
  • Jackwood MW; Poultry Diagnostic and Research Center, Department of Population Health, College of Veterinary Medicine, University of Georgia, Athens GA, United States. Electronic address: mjackwoo@uga.edu.
J Virol Methods ; 276: 113773, 2020 02.
Article en En | MEDLINE | ID: mdl-31712094
Infectious bronchitis (IB) is a highly contagious upper respiratory tract disease of chickens caused by infectious bronchitis virus (IBV), which has various serotypes that do not cross-protect. Vaccine control strategies for this virus are only effective when designed around the currently circulating serotypes. It is essential to not only rapidly detect IBV but also to identify the type of virus causing disease. Six TaqMan™-based quantitative real-time RT-PCR assays (Universal, Ark, Mass, DE/GA98, GA07, GA08) were developed and examined the sensitivity and specificity for each assay. Assays were developed targeting the hypervariable region in the S1 gene subunit. The analytical sensitivity of TaqMan™-based quantitative real-time RT-PCR assays (qRT-PCR) assays was evaluated using synthetic DNA standards that were identical with the target sequence and specificity was further validated using clinical and biological specimens. All developed assays performed equivalently when using synthetic DNA templates as standard material, as it achieved linearity over a 5 log10 dynamic range with a reproducible limit of detection of ≤10 target copies per reaction, with high calculated amplification efficiencies ranging between 90%-115%. Further validation of specificity using clinical and biological specimens was also successful.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Aves / ADN Viral / Infecciones por Coronavirus / Virus de la Bronquitis Infecciosa / Reacción en Cadena en Tiempo Real de la Polimerasa Tipo de estudio: Diagnostic_studies / Guideline / Observational_studies / Prognostic_studies Límite: Animals Idioma: En Revista: J Virol Methods Año: 2020 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Países Bajos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Aves / ADN Viral / Infecciones por Coronavirus / Virus de la Bronquitis Infecciosa / Reacción en Cadena en Tiempo Real de la Polimerasa Tipo de estudio: Diagnostic_studies / Guideline / Observational_studies / Prognostic_studies Límite: Animals Idioma: En Revista: J Virol Methods Año: 2020 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Países Bajos