Brüning A, Bellamy K, Talbot D, Anderson J
Institute for Animal Health, Pirbright Laboratory, Surrey, UK.
J Virol Methods. 1999 Aug;81(1-2):143-54. doi: 10.1016/s0166-0934(99)00068-3.
Rinderpest is a contagious viral disease of cloven-hoofed domestic and wild animals. Eradication of the virus following outbreaks depends on rapid and accurate diagnosis of infection and the implementation of control measures. Reporting and confirmatory diagnosis precede the implementation of control measures. A number of techniques have been used for diagnosis such as agar gel immunodiffusion, enzyme-linked immunosorbent assay (ELISA), molecular biological techniques such as polymerase chain reaction (PCR) and virus isolation in tissue culture. Many of these methods are both time consuming and require skilled personnel. The development of a rapid pen-side test for the detection of rinderpest virus (RPV) antigen in lachrymal fluid of cattle is described using the Clearview chromatographic strip test technology (Unipath, Bedford). Optimum conditions for binding monoclonal antibody to nitrocellulose and latex microspheres were determined and a prototype device was developed. The device detected viral antigen in lachrymal fluids from experimentally and naturally infected cattle and showed no cross-reactivity with other related viruses. A field trial was carried out at the Landhi Cattle Colony (LCC), Pakistan, to assess the performance of the rinderpest test under field conditions. Ninety-seven animals, some of which were showing various clinical signs, at LCC and neighbouring colonies were sampled and tested at the pen-side by Clearview and later by immunocapture ELISA (IC-ELISA) at IAH, Pirbright. Nineteen animals were positive by Clearview and/or IC-ELISA. Seventeen out of 19 rinderpest positive animals were positive by Clearview and 15 out of 19 were positive by IC-ELISA. Reverse transcription polymerase chain reaction (RT-PCR) confirmed the 19 animals to be rinderpest positive. This simple, rapid, specific test allows for the first time, accurate pen-side diagnosis of rinderpest.
牛瘟是一种影响偶蹄目家养和野生动物的传染性病毒病。疫情爆发后根除该病毒取决于对感染的快速准确诊断以及控制措施的实施。报告和确诊诊断先于控制措施的实施。已使用多种技术进行诊断,如琼脂凝胶免疫扩散、酶联免疫吸附测定(ELISA)、分子生物学技术如聚合酶链反应(PCR)以及在组织培养中进行病毒分离。这些方法中的许多既耗时又需要技术熟练的人员。本文描述了利用Clearview色谱试纸条检测技术(Unipath公司,贝德福德)开发一种用于检测牛泪液中牛瘟病毒(RPV)抗原的快速现场检测方法。确定了单克隆抗体与硝酸纤维素和乳胶微球结合的最佳条件,并开发了一种原型装置。该装置检测了来自实验感染和自然感染牛的泪液中的病毒抗原,且与其他相关病毒无交叉反应。在巴基斯坦的兰迪牛群聚居地(LCC)进行了一项现场试验,以评估牛瘟检测方法在现场条件下的性能。对LCC及其周边聚居地的97只动物进行了采样,其中一些动物表现出各种临床症状,在现场通过Clearview方法进行检测,随后在皮布赖特的国际动物卫生研究所通过免疫捕获ELISA(IC - ELISA)进行检测。19只动物通过Clearview和/或IC - ELISA检测呈阳性。19只牛瘟阳性动物中有17只通过Clearview检测呈阳性,19只中有15只通过IC - ELISA检测呈阳性。逆转录聚合酶链反应(RT - PCR)证实这19只动物牛瘟呈阳性。这种简单、快速、特异的检测方法首次实现了牛瘟的准确现场诊断。