Contact Information: Email: email@example.com;Phone: 607-253-3393
Sponsor: USDA-Agriculture Animal and Plant health Inspection Service (APHIS)
Grant Number: 10-9419-0175-CA
Title: Viral Hemorraghic Septicemia Disease Virus Real Time RT-PCR
Annual Direct Cost: $76,516
Project Period: 09/01/10-08/31/11
DESCRIPTION (provided by applicant): The current gold standard for detection of VHSV virus in clinical or nonclinical fish is virus isolation in cell culture followed by virus identification by either fluorescent antibody test or conventional RTPCR assays. This is an expensive, long (14-28 days dependent on the SOP followed), and labor intensive test. Multiple real time RT-PCR (rRT-PCR) assays have been reported for use as a screening assay for detection of VHSV in both clinical and non clinical fish. A screening assay with rapid turnaround time would be useful for surveillance efforts and allow for repeat sampling of populations before the virus has run its course and can no longer be reliably isolated from a recovered population. However, in order to consider utilization of VHSV rRT-PCR assay in surveillance efforts that may be used to support declaring freedom from disease, the assay must be robust and well characterized. This cooperative effort sets out to consider several assays, produce test performance characteristics on multiple platforms for review, and make recommendations regarding utilization of a VHSV rRT-PCR assay.
The objective for this VHSV work plan is to evaluate up to three real time PCR assays and further validate the best fit assay. Specific objectives include:
1. Evaluate up to three real-time PCR assays at the analytical level to determine the best fit assay for detecting all VHSV genotypes and also identify the one with the best sensitivity for VHSV IVb.
2. Evaluate up to two assays at the diagnostic level to determine the best assay for detection of VHSV in samples
3. Determine diagnostic sensitivity and specificity for the best fit assay and characterize robustness and reproducibility on multiple platforms and in different laboratories
This cooperative effort between APHIS and the Cooperator will result in enhanced ability to detect VHSV thus improving the ability to rapidly detect the spread of VHSV throughout the US should it occur, and allowing for effective response to control its spread. Having effective standardized testing capabilities will allow early detection of an incursion of VHSV and thereby decrease response time and facilitate exportation of fish and fish products to US trading partners.