Page 199 - 《水产学报》2026年第3期

3 期 吴海霞，等：草鱼 GCRV-II 抗原检测胶体金试剂盒研发与应用 50 卷

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Development and application of a colloidal gold detection kit for
rapid detection of type II grass carp reovirus

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WU Haixia , FENG Jianhua , LU Yanan , GAO Wa , YAN Hui , XU Dan , JI Xiaohua ,
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LÜ Liqun , XU Zhen , WANG Junya , ZOU Jun 1*
(1. School of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306;
2. Center for Aquatic Breeding Innovation and Future Fisheries Development,
Institute of Hydrobiology, Chinese Academy of Science, Wuhan 430072, China)
Abstract: 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
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reached 1.76×10 copies/μL, and the kit remained stable after storage at 4–25°C 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.
Key words: Ctenopharyngodon idella; GCRV-II; hemorrhagic disease; colloidal gold test kit; rapid detection; monoclonal anti-
body
Corresponding author: ZOU Jun. E-mail: jzou@shou.edu.cn
Funding projects: Major Project for Agricultural and Rural Breeding (2023ZD04065)

中国水产学会主办 sponsored by China Society of Fisheries https://www.china-fishery.cn
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