Kim Hyun-Joo, Nah Jin-Ju, Moon Jin-San, Ko Young-Joon, Yoo Han-Sang, Kweon Chang-Hee
National Veterinary Research and Quarantine Service, Anyang, Gyeonggi 430–824, Republic of Korea.
J Vet Med Sci. 2012 Mar;74(3):321-7. doi: 10.1292/jvms.11-0259. Epub 2011 Oct 20.
Rift Valley fever virus (RVFV) is one of the important emerging viral diseases of serious impact in public health and animal hygiene both in human and animal industries. In this study, we developed a monoclonal antibody-based competitive ELISA for the detection of antibodies to RVFV in goats and cattle. The recombinant N protein of RVFV was expressed in E. coli with a six-histidine tag, and the purified N protein was used for detecting antigen with a competitive monoclonal antibody against RVFV antibodies. The competitive ELISA (C-ELISA) could detect antibodies at 9-11 days after inoculation in goats and cattle with a sensitivity of 94.7% (virus neutralization titer >32) and specificity of 99.7%, respectively. In addition, the C-ELISA did not show any cross-reactivity with positive sera against arboviruses such as Akabane, Aino, Chuzan, Ibaraki and bovine ephemeral fever virus, which are prevalent viral agents in ruminant animals throughout Southeast Asia. The results of the present study indicate that the C-ELISA is a simple, rapid and convenient serodiagnostic method for RVFV in goats and cattle.
裂谷热病毒(RVFV)是一种重要的新出现的病毒性疾病,对人类和动物产业的公共卫生及动物卫生都有严重影响。在本研究中,我们开发了一种基于单克隆抗体的竞争ELISA法,用于检测山羊和牛体内针对RVFV的抗体。RVFV的重组N蛋白在带有六个组氨酸标签的大肠杆菌中表达,纯化后的N蛋白用于与抗RVFV抗体的竞争单克隆抗体一起检测抗原。竞争ELISA(C-ELISA)在山羊和牛接种后9至11天能够检测到抗体,敏感性分别为94.7%(病毒中和效价>32),特异性为99.7%。此外,C-ELISA与针对诸如赤羽病病毒、爱诺病毒、中川病毒、茨城病毒和牛流行热病毒等虫媒病毒的阳性血清没有任何交叉反应,这些病毒是东南亚反刍动物中普遍存在的病毒病原体。本研究结果表明,C-ELISA是一种用于检测山羊和牛体内RVFV的简单、快速且便捷的血清诊断方法。