| 牛口蹄疫病毒VP2结构蛋白抗体间接ELISA方法的建立 |
| |
| 引用本文： | 张润祥,高明春,曲哲会,李爽,曹永生,宋军,宋鸽,刘丹丹,王君伟.牛口蹄疫病毒VP2结构蛋白抗体间接ELISA方法的建立[J].中国预防兽医学报,2010,32(2). |
| |
| 作者姓名： | 张润祥 高明春 曲哲会 李爽 曹永生 宋军 宋鸽 刘丹丹 王君伟 |
| |
| 作者单位： | 1. 东北农业大学,动物医学学院,黑龙江,哈尔滨,150030
2. 信阳农业高等专科学校 动物科学系,河南 信阳,464000 |
| |
| 基金项目： | 国家科技支撑计划，现代农业产业技术体系建设专项资金，黑龙江省科技计划 |
| |
| 摘 要： | 为建立牛口蹄疫(FMD)抗体的检测方法,本研究将口蹄疫病毒(FMDV)的VP2基因,通过pPROEXTM HTb表达载体在大肠杆菌DH5α中表达,获得大小为35ku的重组VP2蛋白(rVP2),western blot证实rVP2可与FMDV5种血清型的牛阳性血清发生特异性反应。以纯化复性的rVP2为抗原建立了FMDVrVP2间接ELISA方法。重复性试验证实批内、批间变异系数均小于10%。特异性交叉试验表明,该抗原不与常见的其他7种牛病阳性血清发生交叉反应。检测非免疫无口蹄疫国家牛阴性血清的特异性为100%;检测感染血清敏感性为97.3%;检测O-AsiaⅠ的二价苗免疫牛血清,与4种商品化试剂盒比较,其符合率分别为69.0%、95.0%、90.4%和86.8%。实验结果表明建立的ELISA方法可以用于口蹄疫感染和免疫抗体检测。
|
| 关 键 词： | 口蹄疫病毒 VP2蛋白 间接ELISA |
Development of VP2 indirect ELISA for detection of cattle antibodies against foot-and-mouth disease virus |
| |
| ZHANG Run-xiang,GAO Ming-chun,Qu Zhe-hui,LI Shuang,CAOYong-sheng,SONG Jun,SONG Ge,LIU Dan-dan,WANG Jun-wei.Development of VP2 indirect ELISA for detection of cattle antibodies against foot-and-mouth disease virus[J].Chinese Journal of Preventive Veterinary Medicine,2010,32(2). |
| |
| Authors: | ZHANG Run-xiang GAO Ming-chun Qu Zhe-hui LI Shuang CAOYong-sheng SONG Jun SONG Ge LIU Dan-dan WANG Jun-wei |
| |
| Institution: | ZHANG Run-xiang1,GAO Ming-chun1,QU Zhe-hui2,LI Shuang1,CAO Yong-sheng1,SONG Jun1,SONG Ge1,LIU Dan-dan1,WANG Jun-wei1 (1.College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China,2. Department of Animal Science,Xinyang Agriculture College,Xinyang 464000,China) |
| |
| Abstract: | The complete gene encoding the structural protein VP2 of FMDV was subcloned into expression vector pPROEX<'TM> HTb and expressed in DH5α cells. Purified VP2 protein reacted positively with serotype-specific cattle sera of 5 serotypes of FMDV (O, A, C, SAT 2 and Asia Ⅰ) by western blot. An indirect ELISA (VP2-ELISA) was developed using purified protein as coating antigen to detect FMDV antibodies in cattle. The assay was highly specific and showed no cross-reaction with the positive sera of other bovine diseases. The sensitivity of the assay was 97.3 % against infected sera. Comparison with four commercial kit showed a coincidence rate of 69.0%, 95.0%, 90.4% and 86.8% against positive cattle serum, respectively. |
| |
| Keywords: | foot-and-mouth disease virus (FMDV) VP2 protein indirect ELISA |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
