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Title: Development and application of an RT-PCR diagnostic test for novel poultry picobirnavirus

Author
item Day, James
item Zsak, Laszlo

Submitted to: World Veterinary Poultry Association
Publication Type: Proceedings
Publication Acceptance Date: 5/2/2011
Publication Date: N/A
Citation: N/A

Interpretive Summary: A recent analysis of the turkey gut virus community in our laboratory revealed numerous un-described enteric viruses that may be associated with disease in the field. This analysis identified the presence of double-stranded RNA viruses (Reoviridae and Picobirnaviruses), and the single-stranded RNA viruses (Caliciviridae, Leviviridae, Picornavirales, and Astroviridae). As a first step in the characterization of these novel enteric viruses, a molecular-based diagnostic test was developed targeting the turkey-origin picobirnavirus present in the turkey gut. Little information exists regarding the picobirnaviruses as a group, and the picobirnaviruses are almost completely un-described in avian species. This molecular assay was validated in our laboratory and was used to test numerous archived enteric samples collected from turkey flocks in the southeastern United States. Data on the incidence of this novel virus in the United States will be presented along with data placing turkey picobirnaviruses among the existing—and still limited—picobirnavirus sequences in national and international databases.

Technical Abstract: A recent metagenomic analysis of the turkey gut ribonucleic acid (RNA) virus community in our laboratory revealed numerous un-described enteric viral sequences. This analysis identified an abundance of sequences from the double-stranded RNA viruses (Reoviridae and Picobirnaviruses), and the single-stranded RNA viruses (Caliciviridae, Leviviridae, Picornavirales, and Astroviridae). As a first step in the characterization of these novel enteric viruses, an reverse transcriptase polymerase chain reaction (RT-PCR) based diagnostic test was developed targeting the turkey-origin picobirnavirus present in the turkey gut. Little information exists regarding the Picobirnaviridae family, and the picobirnaviruses are almost completely un-described in avian species. The novel molecular diagnostic test targets the turkey picobirnavirus RNA-dependent RNA polymerase (RdRp) gene and produces an 1135 base pair amplicon. This RT-PCR test was validated using in vitro transcribed RNA and numerous archived enteric samples collected from turkey flocks in the southeastern United States. Data on the incidence of this novel virus in the United States will be presented along with phylogenetic data placing turkey picobirnavirus among the existing—and still limited—picobirnavirus sequences in the databases.