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使用N蛋白单克隆抗体检测牛瘟病毒。

Detection of rinderpest virus using N-protein monoclonal antibodies.

作者信息

Shah R A, Joseph M C, Butchaiah G, Malik M, Singh R K, Bakshi C S

机构信息

National Biotechnology Centre, Indian Veterinary Research Institute, Izatnagar 243 122, UP, India.

出版信息

Trop Anim Health Prod. 2004 Jan;36(1):11-25. doi: 10.1023/b:trop.0000009527.39602.11.

Abstract

A panel of monoclonal antibodies (mAbs) was generated against the RBOK strain of rinderpest virus (RPV). All of them bound to the N protein of RPV. The antigen capture ELISA using the mAbs could detect the virus in crude viral preparations. The mAb 12BF8.1.1 showed higher reactivity with cell-associated (CA) virus, whereas the mAbs 12AD10.1.1, 12BD7.1.1 and 12DG7.1.1 showed higher reactivity with extracellular virus (hereafter referred to as cell-free (CF) virus). The mAbs 12BF8.1.1 and 12AD10.1.1 could detect the virus in infected Vero cell culture supernatants (CCS) as early as 24 h post-cytopathic effect (CPE) initiation. Detergent treatment (Triton X-100) of RPV preparations enhanced the binding of the mAbs to the virus. All the seven mAbs showed specific fluorescence in virus-infected cell cultures. The immunofluorescence (IFA) using mAbs was found to be more sensitive and reliable than the immunoperoxidase test (IPT) for detection of rinderpest.

摘要

制备了一组针对牛瘟病毒(RPV)RBOK株的单克隆抗体(mAb)。它们均与RPV的N蛋白结合。使用这些mAb的抗原捕获ELISA可检测粗制病毒制剂中的病毒。单克隆抗体12BF8.1.1与细胞相关(CA)病毒反应性较高,而单克隆抗体12AD10.1.1、12BD7.1.1和12DG7.1.1与细胞外病毒(以下称为无细胞(CF)病毒)反应性较高。单克隆抗体12BF8.1.1和12AD10.1.1最早可在细胞病变效应(CPE)开始后24小时检测到感染的Vero细胞培养上清液(CCS)中的病毒。用去污剂(Triton X-100)处理RPV制剂可增强mAb与病毒的结合。所有七种mAb在病毒感染的细胞培养物中均显示出特异性荧光。发现使用mAb的免疫荧光(IFA)在检测牛瘟方面比免疫过氧化物酶试验(IPT)更敏感、更可靠。

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