Department of Fisheries, Faculty of Animal Science and Fisheries, Sari Agricultural Sciences and Natural Resources University, Sari, I.R. Iran.
Department of Scientific Information and Communication, Iranian Fisheries Research Institute (IFSRI), Tehran, I.R. Iran.
J Fish Dis. 2021 Jun;44(6):783-791. doi: 10.1111/jfd.13302. Epub 2021 Feb 1.
A lateral flow immunochromatography strip test, based on antibody-gold nanoparticles specific for nervous necrosis virus (NNV), was developed for rapid, on-site detection of the virus in fish stocks. A monoclonal antibody against NNV was conjugated with colloidal gold as the detector antibody. A rabbit anti-NNV polyclonal antibody and goat anti-mouse IgG antibody were blotted onto the nitrocellulose membrane as the capture antibodies on the test line and control line, respectively. The reaction could be seen by the eye within 15 min and did not cross-react with the other viruses tested. The detection limit of the strip was approximately 10 TCID /ml and had good stability after storage at 4°C for 8 months. When brains of 70 naturally infected golden grey mullet, Chelon aurata, were tested with the strip test, the diagnostic specificity and sensitivity of the test compared to real-time RT-PCR were 100% and 74%, respectively. Therefore, the one-step test strip developed here had high specificity, reproducibility, and stability. This, together with its simplicity to use and rapid detection, without the requirement of sophisticated equipment or specialized skills, makes the strip suitable for pond-side detection of NNV in farmed fish.
一种侧向流动免疫层析条检测法，基于针对神经坏死病毒（NNV）的抗体-胶体金纳米颗粒，被开发用于快速、现场检测鱼类群体中的病毒。一种针对 NNV 的单克隆抗体与胶体金偶联作为检测抗体。兔抗 NNV 多克隆抗体和山羊抗小鼠 IgG 抗体分别被点样到硝酸纤维素膜上作为测试线和控制线的捕获抗体。在 15 分钟内可以通过肉眼观察到反应，并且与测试的其他病毒没有交叉反应。该条带的检测限约为 10 TCID /ml，在 4°C 下储存 8 个月后具有良好的稳定性。当用该条带检测 70 条自然感染的金色灰鲻鱼的脑组织时，与实时 RT-PCR 相比，该测试的诊断特异性和敏感性分别为 100%和 74%。因此，这里开发的一步法测试条具有高特异性、重现性和稳定性。再加上其使用简便、快速检测，不需要复杂的设备或专业技能，使得该条带适合在养殖场池塘边检测养殖鱼类中的 NNV。
