DAO 108:187-199 (2014)  -  DOI: https://doi.org/10.3354/dao02712

Detection of viral hemorrhagic septicemia virus-IVb antibodies in sera of muskellunge Esox masquinongy using competitive ELISA

Elena V. Millard1, Travis O. Brenden2,3, Scott E. LaPatra4, Susan Marcquenski5, Mohamed Faisal1,2,* 

1Department of Pathobiology and Diagnostic Investigation, Michigan State University, East Lansing, Michigan 48824, USA
2Department of Fisheries and Wildlife, Michigan State University, East Lansing, Michigan 48824, USA
3Quantitative Fisheries Center, College of Agriculture and Natural Resources, Michigan State University, East Lansing, Michigan 48824, USA
4Clear Springs Foods, Research Division, Buhl, Idaho 83316, USA
5Wisconsin Department of Natural Resources, Madison, Wisconsin 53707, USA
*Corresponding author:

ABSTRACT: A competitive enzyme-linked immunosorbent assay (cELISA) was developed for the detection of antibodies to viral hemorrhagic septicemia virus genotype IVb (VHSV-IVb) in fish sera. Assay conditions were standardized using known negative and positive muskellunge Esox masquinongy. A positive-negative threshold of 14.6% inhibition was established based on analysis of sera of 60 muskellunge with no previous exposure to VHSV-IVb. The cELISA was then used to investigate immune responses of wild muskellunge sampled from 5 water bodies in Michigan and Wisconsin, USA, between 2005 and 2012. Antibodies were detected in fish from Lake St. Clair, Michigan, and Lower Fox River/Green Bay, Wisconsin. Both water systems were considered enzootic for VHSV-IVb. Additionally, antibodies were detected in muskellunge from Thornapple Lake, a Michigan inland lake previously considered negative for VHSV-IVb based on virus isolation methods. Muskellunge populations from Lake Hudson, Michigan, and Butternut Lake, Wisconsin, lacked evidence of an immune response to VHSV-IVb. When results of the cELISA were compared to the 50% plaque neutralization test for several groups of fish, there was 78.4% agreement between the tests for antibody presence. The cELISA is a rapid and efficient test for the detection of binding antibodies to VHSV-IVb and will be a useful non-lethal tool for monitoring the spread of this serious pathogen.


KEY WORDS: Viral haemorrhagic septicaemia · VHSV · Serology · Surveillance · Immunoassay · Great Lakes · Muskellunge


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Cite this article as: Millard EV, Brenden TO, LaPatra SE, Marcquenski S, Faisal M (2014) Detection of viral hemorrhagic septicemia virus-IVb antibodies in sera of muskellunge Esox masquinongy using competitive ELISA. Dis Aquat Org 108:187-199. https://doi.org/10.3354/dao02712

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