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The purpose of this project request is to continue development of The Nebraska Veterinary Diagnostic Center's (NeVDC) capabilities to function as an integrated diagnostic laboratory resource of the National Animal Health Laboratory Network (NAHLN). As an integrated laboratory, the NeVDC must (1) anticipate the increase in the number of test submissions and (2) strive to improve the efficiency of accurate laboratory testing. The funds requested to support these objectives will be used to verify and validate high throughout screening assays to diagnose influenza virus, pestivirus and bluetongue virus infections.
NON-TECHNICAL SUMMARY: Recent occurrences of influenza virus H1N1infections as well as avian influenza virus infections have demonstrated that regional veterinary diagnostic laboratories must have the technical ability and resources to develop and/or initiate high throughput screening test methods and complementary confirmatory assays for emerging and reemerging infectious diseases. Additionally, high throughput screening test methods and complementary confirmatory assays are essential for testing of all animals in herds in the perimeter of an outbreak of influenza or following introduction of an exotic disease such as classical swine fever. We propose to use endemic ruminant pestiviruses to develop high throughput screening test methods and complementary confirmatory assays that could be applied testing of animals for classical swine fever virus as well as for influenza virus infections. An advantage of PCR assays which will be used is that due to the high test sensitivity, pooled samples can be tested. In this threat environment, diagnostic laboratories must 1) anticipate the increase in the number of test submissions and 2) strive to improve the efficiency of accurate laboratory testing.
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APPROACH: We propose to use endemic ruminant pestiviruses to develop high throughput screening test methods and complementary confirmatory assays that could be applied testing of animals for classical swine fever virus as well as for influenza virus infections. Test methods will be based on pooling and multiplexing for detection of viral RNA in clinical specimens by real-time RT-PCR and incorporate laboratory automation workstations.