传染性法氏囊病毒VP2蛋白的表达和间接.PDF

安徽农业大学学报, 2011, 38(4): 633-636 Journal of Anhui Agricultural University [DOI]CNKI：34-1162/S1624.016 网络出版时间：2011-06-24 16:24:26 [URL]/kcms/detail/34.1162.S1624.016.html 传染性法氏囊病毒VP2蛋白的表达及间接 ELISA抗体检测方法的建立 宋 军，黄成斌，单雪芹，潘 玲* (安徽农业大学动物科技学院，合肥 230036) 摘 要：建立以传染性法氏囊病毒（IBDV ）VP2 蛋白为诊断抗原的传染性法氏囊病（IBD ）血清学检测方法， 研究了 VP2 蛋白的编码基因，并在原核表达系统中表达了 VP2 蛋白。用表达的 VP2 蛋白建立了 IBDV 间接ELISA 检测方法。抗原最佳包被量为每孔 174.67 ng，血清最佳稀释度为 1:40。对采自安徽部分地区的 979 鸡血清样品进 行检测，IBDV 抗体阳性率为 39.73%。结果证实，间接ELISA 法用于 IBD 血清抗体的监测具有良好的特异性，是 一种快速简便的血清学方法，值得在基层推广应用。 关键词：传染性法氏囊病病毒；VP2 蛋白；间接 ELISA；抗体检测 中图分类号：S852.657 文献标识码：A 文章编号：1672−352X (2011)04−0633−04 Establishment of an indirect ELISA method using a infectious bursal disease virus VP2 protein as antigen SONG Jun, HUANG Cheng-bin, SHAN Xue-qin, PAN Ling (School of Animal Science and Technology, Anhui Agricultural University, Hefei 230036) Abstract: The VP2 protein of infectious bursal disease virus(IBDV) is highly conserved and could be used as diagnostic antigen in serological test. In this study, a VP2 protein gene was cloned and expressed in Escherichia coli. The expressed protein was used to establish an indirect ELISA method. The optimal coating concentration of antigen was amounted to 174.67 ng of VP2 protein per well，and the serum sample for testing was diluted to 1:40. A total of 979 serum samples originated from different areas in Anhui province were detected by this indirect ELISA, and the positive rate was 39.73%. The results confirmed that indirect ELISA was specific to monitor se- rum antibody of IBD. This detection method is quick and easy, and it is deserved to be popularized. Key words: infectious bursal disease virus ；VP2 protein ；