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梅迪-维斯纳病诊断的比较研究:免疫扩散试验、酶免疫测定、免疫印迹法

[Comparative studies of maedi-visna diagnosis: immunodiffusion test, enzyme immunoassay, immunoblot].

作者信息

Starick E, Enke K H

机构信息

Bundesforschungsanstalt für Viruskrankheiten der Tiere, Friedrich-Loeffler-Institute, Insel Riems.

出版信息

Berl Munch Tierarztl Wochenschr. 1995 Apr;108(4):138-42.

PMID:7575382
Abstract

The agar gel immunodiffusion test (AGIDT) is applied worldwide for the serological field diagnosis of ovine/caprine lentivirus infections (Maedi-Visna and Caprine Arthritis-Encephalitis). To a lesser extent, enzyme-linked immunosorbent assay (ELISA) systems are in use. The sensitivity and specificity of available ELISA systems is strongly influenced by technical details. We present here an MVV-ELISA of high sensitivity, which is based on an antigen preparation containing a high amount of the major antigenic surface protein gp 135. This result was achieved by a simple and gentle purification method of antigen from whole virus. The test was applied to evaluate the seroprevalence of MVV-antibodies in herds derived from eleven farms located in the North-East of Germany. Furthermore, the antibody kinetics in experimentally infected sheep was investigated. The results were confirmed by internal controls and by immunoblot analysis of antibodies against p 28. The ELISA displayed significant differences between positive and negative results and identified up to 25% more seropositive samples than the AGIDT. Therefore, a final diagnosis of questionable AGIDT results was obtained. The specificity of results was confirmed by immunoblots with selected sera. We recommend to use the ELISA especially for screening of herds and for population controls. In addition, immunoblots are a valuable tool to test individual sera from specified cases.

摘要

琼脂凝胶免疫扩散试验(AGIDT)在全球范围内用于绵羊/山羊慢病毒感染(梅迪 - 维斯纳病和山羊关节炎 - 脑炎)的血清学现场诊断。在较小程度上,酶联免疫吸附测定(ELISA)系统也在使用。现有ELISA系统的敏感性和特异性受技术细节的强烈影响。我们在此展示一种高灵敏度的梅迪 - 维斯纳病ELISA(MVV - ELISA),它基于一种含有大量主要抗原表面蛋白gp 135的抗原制剂。这一结果是通过一种从全病毒中简单温和的抗原纯化方法实现的。该检测方法用于评估德国东北部11个农场的畜群中MVV抗体的血清流行率。此外,还研究了实验感染绵羊的抗体动力学。结果通过内部对照以及针对p 28抗体的免疫印迹分析得到证实。该ELISA在阳性和阴性结果之间显示出显著差异,并且比AGIDT多识别出高达25%的血清阳性样本。因此,对AGIDT可疑结果做出了最终诊断。结果的特异性通过对选定血清的免疫印迹得到证实。我们建议特别使用ELISA进行畜群筛查和群体监测。此外,免疫印迹是检测特定病例个体血清的有价值工具。

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