Department of Neurovirology, National Institute of Mental health and Neurosciences, Bangalore 560029, India.
Virol Sin. 2012 Oct;27(5):299-302. doi: 10.1007/s12250-012-3265-6. Epub 2012 Oct 11.
Presently the gold standard diagnostic technique for rabies is the direct immunofluorescence assay (dFA) which is very expensive and requires a high level of expertise. There is a need for more economical and user friendly tests, particularly for use in developing countries. We have established one such test called the direct rapid immunohistochemical test (dRIT) for diagnosis of rabies using brain tissue. The test is based on capture of rabies nucleoprotein (N) antigen in brain smears using a cocktail of biotinylated monoclonal antibodies specific for the N protein and color development by streptavidin peroxidase-amino ethyl carbazole and counter staining with haematoxollin. The test was done in parallel with standard FAT dFA using 400 brain samples from different animals and humans. The rabies virus N protein appears under light microscope as reddish brown particles against a light blue background. There was 100 % correlation between the results obtained by the two tests. Also, interpretation of results by dRIT was easier and only required a light microscope. To conclude, this newly developed dRIT technique promises to be a simple, cost effective diagnostic tool for rabies and will have applicability in field conditions prevalent in developing countries.
目前,狂犬病的金标准诊断技术是直接免疫荧光检测法(dFA),该方法非常昂贵,需要高度专业知识。需要更经济和用户友好的测试方法,特别是在发展中国家。我们已经开发了一种称为直接快速免疫组织化学检测法(dRIT)的测试方法,用于使用脑组织诊断狂犬病。该测试基于使用针对 N 蛋白的生物素化单克隆抗体混合物捕获脑涂片上的狂犬病核蛋白(N)抗原,并通过链霉亲和素过氧化物酶-氨基乙基咔唑显色,并用苏木精复染。该测试与标准 FAT dFA 平行进行,使用来自不同动物和人类的 400 个脑组织样本。狂犬病病毒 N 蛋白在光镜下呈现出淡蓝色背景上的红棕色颗粒。两种测试的结果具有 100%的相关性。此外,dRIT 的结果解释更简单,仅需要一台光学显微镜。总之,这种新开发的 dRIT 技术有望成为一种简单、经济有效的狂犬病诊断工具,并将在发展中国家普遍存在的现场条件下具有适用性。