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Title: A multiplex RT-PCR test for the differential identification of turkey astrovirus type-1, turkey astrovirus type-2, chicken astrovirus, avian nephritis virus and avian rotavirus

Author
item Day, James
item Pantin Jackwood, Mary
item Spackman, Erica

Submitted to: Avian Diseases
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/26/2007
Publication Date: 9/21/2007
Citation: Day, J.M., Pantin Jackwood, M.J., Spackman, E. 2007. A multiplex RT-PCR test for the differential identification of turkey astrovirus type-1, turkey astrovirus type-2, chicken astrovirus, avian nephritis virus and avian rotavirus. Avian Diseases. 51:681-684.

Interpretive Summary: Viral enteric diseases cause substantial economic loss to the US poultry industry. Viruses from two distinct families, avian astroviruses and avian rotaviruses, are among the most commonly detected viruses found in specimens from chickens and turkeys with enteric disease. Although rotavirus can be easily distinguished from astrovirus, within the astrovirus group there are four types which previously could only be distinguished by gene sequencing. Gene sequencing is time consuming, expensive and is therefore not within the capabilities of most diagnostic labs. This manuscript reports the development of RT-PCR tests for the simultaneous detection and differentiation of four avian astrovirus types as well as the detection avian rotaviruses from feces and intestinal samples from poultry. The test has two variants; one for chicken-origin specimens and one for turkey-origin specimens. The sensitivity and sensitivity of the test was also evaluated as part of test development.

Technical Abstract: Viral enteric diseases cause substantial economic loss to the US poultry industry because they lead to decreased weight gain, increased morbidity, increased mortality, and increased production costs from poor feed conversions and increased use of therapeutic anti-microbial treatments. Astroviruses and rotaviruses are among the most commonly detected viruses found in chicken and turkey intestinal samples. Two multiplex RT-PCR tests for the simultaneous detection and differentiation of four avian astrovirus types and avian rotaviruses from feces and intestinal samples from poultry were developed. The test has two variants; one for chicken-origin specimens and one for turkey-origin specimens. The multiplex RT-PCR for chicken-origin specimens differentially detects Avian Nephritis Virus (ANV), chicken astrovirus and avian rotavirus. The multiplex RT-PCR for turkey-origin specimens differentially detects turkey astrovirus type-1, turkey astrovirus type-2, ANV and avian rotavirus. The analytical sensitivity of the assay was determined to for each virus individually with in vitro transcribed RNA and were shown to be equivalent to similar tests. The specificity of each assay was confirmed by sequencing of the amplicons from field samples collected during a survey of enteric viruses circulating in poultry in the United States and by testing for cross reaction with numerous common viruses of poultry. This test will save time and cost since gene sequencing will no longer be necessary to differentiate among these viruses.