National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai, China.
Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding, Shanghai Ocean University, Shanghai, China.
J Fish Dis. 2023 Oct;46(10):1065-1071. doi: 10.1111/jfd.13825. Epub 2023 Jul 6.
Cyprinid herpesvirus 3 (CyHV-3) is the main pathogen of koi herpesvirus disease (KHVD), which has caused serious damage to the ornamental and food-producing carp industry. Effective and rapid on-site detection methods are needed for early diagnosis of CyHV-3. A lateral flow immuno-chromatographic assay (LFIA) using two specific anti-CyHV-3 monoclonal antibodies has been developed and validated for on-site detection of CyHV-3. MAb 3C9 was used to bio-conjugate CyHV-3 antigen with colloidal gold, and MAb 2A8 was used to capture antigen bound colloidal gold on the test line. The control line was lined with goat anti-mouse IgG to capture unbound colloidal gold to validate performance. The test results can be viewed within 10 min after putting the strip into CyHV-3 virus infection fluid. The lowest limit of detection for the LFIA test was found to be 1.5 × 10 copies/μL and it showed no cross-reactivity with other fish viral pathogens. The specificity of the strip was 100% when spleen and kidney tissues of CyHV-3-infected and healthy koi were validated at the field level. The LFIA strip will be an effective device for the early detection of CyHV-3 in the future.
鲤鱼疱疹病毒 3（CyHV-3）是锦鲤疱疹病毒病（KHVD）的主要病原体，对观赏鱼和食用鲤鱼产业造成了严重破坏。需要有效的快速现场检测方法来对 CyHV-3 进行早期诊断。已经开发并验证了一种使用两种特异性抗 CyHV-3 单克隆抗体的侧向流动免疫层析测定法（LFIA），用于现场检测 CyHV-3。MAb 3C9 被用于将 CyHV-3 抗原与胶体金生物偶联，MAb 2A8 被用于捕获测试线上结合的胶体金抗原。控制线用山羊抗小鼠 IgG 包被，以捕获未结合的胶体金来验证性能。将条放入 CyHV-3 病毒感染液后 10 分钟内即可查看测试结果。该 LFIA 测试的最低检测限为 1.5×10 拷贝/μL，与其他鱼类病毒病原体无交叉反应。在现场水平验证 CyHV-3 感染和健康锦鲤的脾脏和肾脏组织时，该条的特异性为 100%。该 LFIA 条将是未来早期检测 CyHV-3 的有效设备。
