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The Use of a One‐Step Real‐Time Reverse Transcription Polymerase Chain Reaction (rRT‐PCR) for the Surveillance of Viral Hemorrhagic Septicemia Virus (VHSV) in Minnesota
Author(s) -
Phelps Nicholas B. D.,
Patnayak Devi P.,
Jiang Yin,
Goyal Sagar M.
Publication year - 2012
Publication title -
journal of aquatic animal health
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 52
eISSN - 1548-8667
pISSN - 0899-7659
DOI - 10.1080/08997659.2012.711268
Subject(s) - virology , biology , virus , reverse transcription polymerase chain reaction , outbreak , polymerase chain reaction , real time polymerase chain reaction , reverse transcriptase , isolation (microbiology) , virulence , microbiology and biotechnology , gene , genetics , messenger rna
Viral hemorrhagic septicemia virus (VHSV) is a highly contagious and pathogenic virus of fish. The virus infects more than 70 fish species worldwide, in both fresh and salt water. A new viral strain (VHSV‐IVb) has proven both virulent and persistent, spreading throughout the Great Lakes of North America and to inland water bodies in the region. To better understand the geographic distribution of the virus, we used a modified real‐time reverse transcription polymerase chain reaction (rRT‐PCR) assay for high‐throughput testing of fish for VHSV. The assay was shown to be twice as sensitive as the gold standard, virus isolation, and did not cross react with other viruses found in fish. In addition, the diagnostic turnaround time was reduced from 28 to 30 d for virus isolation to 2–4 d for rRT‐PCR. To demonstrate the usefulness of the rRT‐PCR assay, 115 high‐priority water bodies in Minnesota were tested by both methods from April 2010 to June 2011. All survey sites tested negative for VHSV by both methods. The survey results have informed fisheries managers on the absence of VHSV in Minnesota and have better prepared them for the eventual arrival of the disease. In addition, the results demonstrate the value of this rRT‐PCR as a surveillance tool to rapidly identify an outbreak so that it can be controlled in a timely manner. Received April 24, 2012; accepted July 2, 2012