Serodiagnosis of grass carp reovirus infection in grass carp Ctenopharyngodon idella by a novel Western blot technique.
Frequent outbreaks of grass carp hemorrhagic disease, caused by grass carp reovirus (GCRV) infection, pose as serious threats to the production of grass carp Ctenopharyngodon idella. Although various nucleic acids-based diagnostic methods have been shown effective, lack of commercial monoclonal antibody against grass carp IgM has impeded the development of any reliable immunoassays in detection of GCRV infection. The present study describes the preparation and screening of monoclonal antibodies against the constant region of grass carp IgM protein, and the development of a Western blot (WB) protocol for the specific detection of antibodies against outer capsid VP7 protein of GCRV that serves as antibody-capture antigen in the immunoassay. In comparison to a conventional RT-PCR method, validity of the WB is further demonstrated by testing on clinical fish serum samples collected from a grass carp farm in Jiangxi Province during disease pandemic in 2011. In conclusion, the WB technique established in this study could be employed for specific serodiagnosis of GCRV infection.
National Pathogen Collection Center for Aquatic Animals, Key Laboratory of Freshwater Fishery Germplasm Resources, Shanghai Ocean University, 201306 Shanghai, China.
Yongxing
Y
He
Yousheng
Y
Jiang
Liqun
L
Lu
D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000097:blood*; D015153:Blotting, Western / Q000379:methods*; D002347:Carps; D005393:Fish Diseases / Q000175:diagnosis* / Q000821:virology; D007075:Immunoglobulin M / Q000097:blood; D012087:Reoviridae / Q000276:immunology / Q000302:isolation & purification*; D012088:Reoviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods; D014730:Veterinary Medicine / Q000379:methods*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D007075:Immunoglobulin M
Grass carp reovirus; IgM; Monoclonal antibody; VP7; Western blot