Li Qingmei, Wang Li, Sun Yaning, Liu Jinling, Ma Fansu, Yang Jifei, Zhao Dong, Zhang Yuhang, Luo Jun, Guo Junqing, Deng Ruiguang, Zhang Gaiping
Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou, China (Li, Wang, Sun, Liu, Ma, Yang, Zhao, Luo, Guo, Deng).
College of Animal Husbandry and Veterinary Sciences, Henan Agricultural University, Zhengzhou, China (Y Zhang, G Zhang).
J Vet Diagn Invest. 2019 May;31(3):475-480. doi: 10.1177/1040638719837320. Epub 2019 Apr 11.
We evaluated an immunochromatographic strip for the detection of avian avulavirus 1 (Newcastle disease virus, NDV) based on a high-affinity monoclonal antibody (mAb) that specifically recognizes the hemagglutinin-neuraminidase (HN) protein. The anti-HN mAb was labeled with colloidal gold as the detector. A chicken anti-NDV polyclonal antibody and staphylococcal protein A (SPA) were blotted on the nitrocellulose membrane for the test and control lines, respectively. The strip specifically recognized the NDV antigen with no cross-reactivity to other viruses that were examined. Furthermore, it specifically recognized a variety of NDV isolates, including virulent and attenuated strains. These results were confirmed using hemagglutination (HA) and RT-PCR assays. The NDV detection strip detected 10 EID viruses/0.1 mL in the NDV-infected sample, which is comparable to the classical HA test (10 EID/0.1 mL). Following experimental infection, NDV was detected using the detection strip in infected tissues as early as 36 h after experimental infection and prior to development of clinical signs and appearance of gross anatomic lesions. The diagnostic sensitivity and specificity of the NDV detection strip for NDV infection were 83.3% and 100%, respectively, as confirmed by RT-PCR.
我们评估了一种基于高亲和力单克隆抗体（mAb）的免疫层析试纸条，用于检测禽1型副粘病毒（新城疫病毒，NDV），该单克隆抗体可特异性识别血凝素神经氨酸酶（HN）蛋白。抗HN单克隆抗体用胶体金标记作为检测剂。鸡抗NDV多克隆抗体和葡萄球菌蛋白A（SPA）分别印迹在硝酸纤维素膜上作为检测线和对照线。该试纸条能特异性识别NDV抗原，与所检测的其他病毒无交叉反应。此外，它能特异性识别多种NDV分离株，包括强毒株和弱毒株。这些结果通过血凝试验（HA）和逆转录-聚合酶链反应（RT-PCR）分析得到证实。NDV检测试纸条在NDV感染样本中能检测到10个鸡胚感染剂量（EID）病毒/0.1 mL，这与经典的HA试验（10 EID/0.1 mL）相当。实验感染后，早在实验感染后36小时，在出现临床症状和大体解剖病变之前，就可以使用检测试纸条在感染组织中检测到NDV。经RT-PCR证实，NDV检测试纸条对NDV感染的诊断敏感性和特异性分别为83.3%和100%。
