title: Detection of grass carp reovirus (GCRV) with monoclonal antibodies.
abstract: Grass carp reovirus (GCRV) is a pathogen that causes hemorrhagic disease of grass carp. It is the most serious infectious disease of carp and causes serious losses of fingerlings of grass carp and black carp. In this study, a recombinant VP4, one of the viral core proteins, was constructed with a histidine tag and expressed at a high level in E. coli, and the expressed protein was mainly found in the form of inclusion bodies. The expressed VP4 protein was recognized by an anti-His-tag monoclonal antibody and goat anti-GCRV serum. Four monoclonal antibodies (16B7, 39E12, 13C3 and 14D1) against the recombinant VP4 protein were produced. These MAbs did not react with any of the tested viruses or fish cells lines in the ELISA tests except GCRV. In western blotting analysis, a protein band was observed when the recombinant VP4 protein of GCRV was used as an antigen, but a 68-kDa band was observed when natural capsid proteins of GCRV were used as antigens. Furthermore, a sandwich ELISA was developed for detection of GCRV. The detection limit of the test was 105 TCID50 of GCRV per mL.
authors.forename: Jing
authors.initials: J
authors.lastname: Hongli
authors.forename: Zhang
authors.initials: Z
authors.lastname: Lifeng
authors.forename: Fang
authors.initials: F
authors.lastname: Zhenzhen
authors.forename: Xu
authors.initials: X
authors.lastname: Lipu
authors.forename: Zhang
authors.initials: Z
authors.lastname: Min
authors.forename: Wang
authors.initials: W
authors.lastname: Na
authors.forename: Jiang
authors.initials: J
authors.lastname: Yulin
authors.forename: Lin
authors.initials: L
authors.lastname: Xiangmei
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000302:isolation & purification; D015153:Blotting, Western / Q000379:methods; D002347:Carps / Q000821:virology; D003955:Diagnostic Tests, Routine / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005393:Fish Diseases / Q000821:virology*; D012087:Reoviridae / Q000302:isolation & purification*; D012088:Reoviridae Infections / Q000175:diagnosis* / Q000821:virology; D012680:Sensitivity and Specificity; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral