title: Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus.
abstract: The infectious bursal disease virus (IBDV) is a major health threat to the world's poultry industry despite intensive controls including proper biosafety practices and vaccination. IBDV (Avibirnavirus, Birnaviridae) is a non-enveloped virus with a bisegmented double-stranded RNA genome. The virus is traditionally classified into classic, variant and very virulent strains, each with different epidemiological relevance and clinical implications. Recently, a novel worldwide spread genetic lineage was described and denoted as distinct (d) IBDV. Here, we report the development and validation of a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay for the specific detection of dIBDVs in the global poultry industry. The assay employs a TaqMan-MGB probe that hybridizes with a unique molecular signature of dIBDV. The assay successfully detected all the assessed strains belonging to the dIBDV genetic lineage, showing high specificity and absence of cross-reactivity with non-dIBDVs, IBDV-negative samples and other common avian viruses. Using serial dilutions of in vitro-transcribed RNA we obtained acceptable PCR efficiencies and determination coefficients, and relatively small intra- and inter-assay variability. The assay demonstrated a wide dynamic range between 103and 108RNA copies/reaction. This rapid, specific and quantitative assay is expected to improve IBDV surveillance and control worldwide and to increase our understanding of the molecular epidemiology of this economically detrimental poultry pathogen.
authors.affiliation: a Sección Genética Evolutiva, Departamento de Biología Animal , Instituto de Biología, Facultad de Ciencias, Universidad de la República , Montevideo , Uruguay.
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Tomás
authors.affiliation: a Sección Genética Evolutiva, Departamento de Biología Animal , Instituto de Biología, Facultad de Ciencias, Universidad de la República , Montevideo , Uruguay.
authors.forename: Martín
authors.initials: M
authors.lastname: Hernández
authors.affiliation: a Sección Genética Evolutiva, Departamento de Biología Animal , Instituto de Biología, Facultad de Ciencias, Universidad de la República , Montevideo , Uruguay.
authors.forename: Ana
authors.initials: A
authors.lastname: Marandino
authors.affiliation: a Sección Genética Evolutiva, Departamento de Biología Animal , Instituto de Biología, Facultad de Ciencias, Universidad de la República , Montevideo , Uruguay.
authors.forename: Claudia
authors.initials: C
authors.lastname: Techera
authors.affiliation: a Sección Genética Evolutiva, Departamento de Biología Animal , Instituto de Biología, Facultad de Ciencias, Universidad de la República , Montevideo , Uruguay.
authors.forename: Sofia
authors.initials: S
authors.lastname: Grecco
authors.affiliation: a Sección Genética Evolutiva, Departamento de Biología Animal , Instituto de Biología, Facultad de Ciencias, Universidad de la República , Montevideo , Uruguay.
authors.forename: Diego
authors.initials: D
authors.lastname: Hernández
authors.affiliation: b Poultry Research and Diagnostic Laboratory, College of Veterinary Medicine , Mississippi State University , Pearl , MS , USA.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Banda
authors.affiliation: a Sección Genética Evolutiva, Departamento de Biología Animal , Instituto de Biología, Facultad de Ciencias, Universidad de la República , Montevideo , Uruguay.
authors.forename: Yanina
authors.initials: Y
authors.lastname: Panzera
authors.affiliation: a Sección Genética Evolutiva, Departamento de Biología Animal , Instituto de Biología, Facultad de Ciencias, Universidad de la República , Montevideo , Uruguay.
authors.forename: Ruben
authors.initials: R
authors.lastname: Pérez
mesh_terms: D000818:Animals; D018175:Birnaviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D002060:Bursa of Fabricius / Q000821:virology; D002645:Chickens / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes; D007243:Infectious bursal disease virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology; D012330:RNA, Double-Stranded / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D015678:Viral Structural Proteins / Q000235:genetics*
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D012330:RNA, Double-Stranded; C082513:VP2 protein, infectious bursal disease virus; D015678:Viral Structural Proteins
keywords: IBD; IBDV; RT-qPCR; TaqMan-MGB probe; distinct IBDV