Project number: 2001-626
Project Status:
Completed
Budget expenditure: $185,902.00
Principal Investigator: Nicholas J. Moody
Organisation: Department of Agriculture and Fisheries EcoScience Precinct
Project start/end date: 27 Feb 2002 - 30 Jun 2005
Contact:
FRDC

Need

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.

Immunodiagnostics

- 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.

Objectives

1. To optimise and validate a sensitive and specific Nested RT-PCR test for the detection and identification of endemic and exotic nodaviruses from a range of samples and fish species.
2. To establish a cell line which can be used for the isolation, amplification and titration of endemic and exotic nodaviruses from a range fish species.
3. To produce immunodiagnostic tests, capable of localising endemic and exotic nodaviruses in fish tissues and cell cultures.
4. To distribute the above technology and protocols to laboratories as soon as optimisation and validation are complete.
5. To produce an Australian and New Zealand Standard Diagnostic Procedure for the detection of Nodavirus.

Final report

Related research

Industry
Industry
Industry
PROJECT NUMBER • 2018-140
PROJECT STATUS:
CURRENT

Strengthening the ABFA Quality Framework

1. Industry quality standards for farmed Australian Barramundi are to be enhanced and implemented
ORGANISATION:
Australian Barramundi Farmers Association (ABFA)