草鱼GCRV-Ⅱ抗原检测胶体金试剂盒研发与应用OA

Grass carp(Ctenopharyngodon idella)hemorrhagic disease,caused by grass carp reovirus type Ⅱ(GCRV-Ⅱ),is the most devastating viral disease affecting grass carp aquaculture in China.GCRV-Ⅱ is highly virulent and can rapidly spread,leading to substantial economic losses annually.Current detection of GCRV still heavily relies on laboratory-based molecular techniques,which are complex and fail to meet the urgent need for on-site rapid diagnosis.To address this,this study aims to develop a rapid detection kit for GCRV-Ⅱ based on colloidal gold immunochromatography.Two monoclonal antibodies,VP35#8 and VP35#18,specifically recognizing two distinct epitopes of the outer capsid protein VP35 of GCRV-Ⅱ,were util-ized.The VP35#8 antibody was conjugated with 20 nm colloidal gold particles to prepare the gold-labeled pad,while the VP35#18 antibody and goat anti-mouse IgG were immobilized on the test line(T-line)and control line(C-line)of the nitrocel-lulose membrane,respectively,to assemble the test strip.The results demonstrated that the kit could successfully detect GCRV-Ⅱ from diseased C.idella and Gobiocypris rarus within 10 minutes.It showed excellent specificity with no cross-reactivity against Spring Viraemia of Carp Virus(SVCV),Cyprinid Herpesvirus 3(CyHV-3),and GCRV-Ⅰ.The detection sensitivity reached 1.76×102 copies/μL,and the kit remained stable after storage at 4-25℃ for 12 months.The results were consistent with that obtained by the national standard method(GB/T 36 190-2018).In summary,this study successfully developed a user-friendly,rapid,and reliable on-site detection tool for GCRV-Ⅱ,providing solid technical support for the early diagnosis and control of C.idella hemorrhagic disease,which is of great significance for safeguarding the sustainability of the aquaculture industry.