Centre for Animal Disease Research and Diagnosis (CADRAD), Indian Veterinary Research Institute, Izatnagar, 243122, U.P., India.
Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, 243122, U.P., India.
J Virol Methods. 2018 Jul;257:69-72. doi: 10.1016/j.jviromet.2018.04.007. Epub 2018 Apr 13.
Avian reovirus (ARV) causes significant economic losses to the poultry industry worldwide. The ARV proteins fall into three different classes based on their sizes:λ (large); μ (medium) and σ (small). σB, an outer capsid protein of the ARV contains group specific neutralizing epitopes and induces strong immune response in naturally infected chickens. This study describes the development of a rapid dot-enzyme linked immunosorbent assay (dot-ELISA) using recombinant σB protein antigen of 54 kDa (approx). The assay is rapid (4-5 h) and results can be read by the naked eye. Sixteen ARV positive serum samples (group A) produced strong reaction in the dot-ELISA while twenty of the ARV negative serum samples (group B) collected from SPF chickens showed no reaction. Seventy six randomly collected serum samples were tested with a commercial indirect ELISA kit and the in-house developed dot-ELISA. A total of sixty eight serum samples were found to be positive by indirect ELISA and sixty five serum samples were found to be positive by dot-ELISA. Therefore, using the commercial ELISA as the reference test, the dot-ELISA had a diagnostic sensitivity of 83.8% and specificity of 88.6%. This dot-ELISA can be used as a simple, reliable and inexpensive alternative to commercial ELISA kits for serodiagnosis of ARV where the facilities for standard ELISA are not available.
禽呼肠孤病毒（ARV）会给全球的家禽业造成巨大的经济损失。根据大小，ARV 蛋白分为三类：λ（大）、μ（中）和 σ（小）。σB 是 ARV 的外壳蛋白，含有具有组特异性的中和表位，能在自然感染的鸡中引发强烈的免疫反应。本研究描述了使用 54kDa（约）重组 σB 蛋白抗原建立快速斑点酶联免疫吸附试验（dot-ELISA）的方法。该检测方法快速（4-5 小时），结果可肉眼判读。16 份 ARV 阳性血清样本（A 组）在 dot-ELISA 中产生强烈反应，而 20 份来自 SPF 鸡的 ARV 阴性血清样本（B 组）则无反应。76 份随机采集的血清样本用商业间接 ELISA 试剂盒和内部开发的 dot-ELISA 进行了检测。共有 68 份血清样本用间接 ELISA 检测为阳性，65 份血清样本用 dot-ELISA 检测为阳性。因此，以商业 ELISA 作为参考检测方法，dot-ELISA 的诊断灵敏度为 83.8%，特异性为 88.6%。这种 dot-ELISA 可作为一种简单、可靠和经济的替代方法，用于无法进行标准 ELISA 的 ARV 血清学诊断。
