建立一种间接的ELISA方法来检测基于N蛋白的牛冠状病毒抗体
Qiang Liu1, Xiaoxia Niu1, Lingling Jiang1
1Key Lab of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, School of Life Sciences, Ningxia University, Yinchuan, China.
Frontiers in veterinary science
|February 20, 2025
概括
一种新的间接酶相关免疫吸收试验 (iELISA) 能够有效地检测牛群新冠病毒 (BCoV) 抗体. 这种经济高效的方法为BCoV提供了精确的诊断,有助于疾病管理和流行病学研究.
科学领域:
- 兽医病毒学 兽医病毒学
- 免疫诊断 免疫诊断 免疫诊断 免疫诊断
- 分子生物学分子生物学
背景情况:
- 牛牛冠状病毒 (BCoV) 由于新生儿小牛腹和呼吸道疾病等疾病,在牛畜行业造成了重大经济损失.
- 准确和快速检测BCoV对于有效的疾病管理和控制战略至关重要.
- 目前的诊断方法需要在成本效益和精度方面进行改进.
研究的目的:
- 开发和验证一种新的间接与酶相关的免疫吸收试验 (iELISA) 用于检测针对牛冠状病毒 (BCoV) 的抗体.
- 用BCoV的重组核体 (N) 蛋白用于抗体检测.
- 在临床和流行病学环境中为BCoV建立一个具有成本效益和精确的诊断工具.
主要方法:
- 重组BCoV核体 (N) 蛋白在CHO细胞中得到表达.
- 子对N蛋白的多克隆抗体 (pAb) 通过免疫产生.
- 对抗原和抗体度进行了间接ELISA (iELISA) 优化,确定了0.986.6的切割值.
- 通过对其他牛病毒 (BHV-1,BVDV,BRV,BRSV) 的测试证实了特异性.
主要成果:
- 开发的iELISA表现出高灵敏度,最大抗体稀释值为2^18.
- 该试验显示出优异的特异性,没有针对BHV-1,BVDV,BRV和BRSV阳性血清的交叉反应.
- 与使用58个牛血清样本的商业套件进行比较,得出了高达94.83%的一致率.
结论:
- 已经成功开发了一种精确且具有成本效益的iELISA方法来检测BCoV抗体.
- 基于N蛋白的iELISA作为一种有价值的诊断工具,用于在牛群中检测BCoV.
- 这种测试适用于BCoV感染的临床诊断和流行病学研究.
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