Validate a sensitive and specific Nested RT-PCR test.
- There is a need to ensure the Nested RT-PCR test developed at OVL will detect the range of endemic nodaviruses from a variety of finfish species. It is also important to ensure the test will detect exotic nodavirus isolates for use in the event of an exotic nodavirus incursion.
- Standardisation of reagents and sample collection and preservation protocols will enable consistency of test methods between laboratories.
- A test is required to screen broodstock samples to eliminate nodavirus-positive carrier fish from production facilities. Due to the small sample sizes obtainable, the Nested RT-PCR test is the best test option.
Establishment of a cell line.
- Cell culture is considered the gold standard for virus detection. The cell culture will enable cost-effective screening of larvae for sale or release and to develop management procedures.
- Cell culture is more tolerant to sample degradation than other detection methods and allows testing of material that is unsuitable for use in other tests.
- The vast majority of cell lines currently available have been produced from temperate species. Availability of cell lines, from tropical fish species, is required.
- FAT tests can confirm the identity of viruses isolated in cell culture during diagnostic or surveillance activities. Their use is critical for the identification of viruses.
- Immunohistochemistry/immunofluorescence is a sensitive and specific test that can be used on fixed material. It is not always possible to obtain fresh samples and confirmation of nodavirus in tissue sections can be required. Immunohistochemistry is also a tool to identify the tissues targeted by nodavirus.