Development of a quantitative Reverse Transcription PCR assay for detection of IHNV and determination of optimal sampling protocols

P-07-04-011

Description

The source of the IHNV introduction to farmed salmon is unknown, but epidemiological investigations have identified sockeye salmon and herring as likely sources. Due to the potential devastating effect of IHNV on the economic sustainability of the BC salmon aquaculture industry, companies have developed biosecurity action plans for viral containment in the event of another outbreak. However, effectiveness of any containment plan depends on rapid diagnosis of the index case. Therefore, rapid and accurate diagnosis of IHNV is essential. The traditional method of diagnosing IHNV was through recognizing necrosis of cells grown in tissue culture - a technique requiring between 5 and 21 days for confirmation of virus.

Quantitative PCR (QPCR) is rapidly replacing more traditional methodologies as a diagnostic test. QPCR offers many advantages over other diagnostic techniques including a fast turn-around time as well as reduced frequency of false positives, increased sensitivity, low requirements for tissue and high sample through-out. This technology can also be employed in the detection of IHNV but must include an additional step owing to the fact that the genome of IHNV is composed of single-stranded, negative-sense RNA. Therefore, a reverse transcription step is required to convert genomic and mRNA to cDNA. This proposal seeks to develop a qRT-PCR assay for detection of IHNV.

Program Name

Aquaculture Collaborative Research and Development Program (ACRDP)

Year(s)

2007 - 2009

Ecoregion(s)

National

Principal Investigator(s)

Kyle Garver
Email: garverk@pac.dfo-mpo.gc.ca