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| 检测牛边缘无浆体抗体的竞争抑制ELISA方法的建立 | |
| 引用本文： | 倪宏波,王冰,姜海芳,周玉龙,王春仁,宫大庆,钱爱东.检测牛边缘无浆体抗体的竞争抑制ELISA方法的建立[J].中国预防兽医学报,2011,33(4). |
| 作者姓名： | 倪宏波 王冰 姜海芳 周玉龙 王春仁 宫大庆 钱爱东 |
| 作者单位： | 1. 吉林农业大学动物科技学院,吉林长春130118;黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 2. 黑龙江农垦职业学院食品工程系,黑龙江哈尔滨,150025 3. 黑龙江八一农垦大学动物科技学院,黑龙江大庆,163319 4. 吉林农业大学动物科技学院,吉林长春,130118 |
| 基金项目： | 吉林省科技发展计划项目，黑龙江省科技厅实验动物青年基金项目 |
| 摘 要： | 为建立检测牛边缘无浆体(Anaplasma marginale)抗体的方法,本研究以牛A.marginale膜表面重组MSP5蛋白作为包被抗原,抗MSP5单克隆抗体(MAb)作为竞争抗体,建立一种用于检测牛A marginale抗体的重组MSP5蛋白竞争抑制ELISA(CI-ELISA)方法.经优化确定CI-ELISA的最佳反应条件为:抗原包被浓度为2μg/孔,封闭液为2%脱脂乳,MAb的稀释度为1:400,酶标二抗的稀释度为1:1000,阴性和阳性血清临界值分别为33%和40%;该方法具有良好的特异性和重复性;2 348份临床血清样品的检测结果表明,217份为阳性,阳性率为9.2%,与IDEXXA marginale抗体检测试剂盒的阳性符合率为95.3%,阴性符合率为100%.本实验建立的ELISA方法具有较高的特异性和重复性,可用于流行病学调查研究. |
| 关 键 词： | 牛边缘无浆体 竞争抑制ELISA 重组MSP5蛋白 流行病学调查 |
Establishment of a competitive inhibition ELISA for detection of antibodies against bovine Anaplasma marginale |
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| NI Hong-bo,WANG Bing,JIANG Hai-fang,ZHOU Yu-long,WANG Chun-ren,GONG Da-qing,QIAN Ai-dong.Establishment of a competitive inhibition ELISA for detection of antibodies against bovine Anaplasma marginale[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(4). | |
| Authors: | NI Hong-bo WANG Bing JIANG Hai-fang ZHOU Yu-long WANG Chun-ren GONG Da-qing QIAN Ai-dong |
| Institution: | NI Hong-bo1,3,WANG Bing2,JIANG Hai-fang3,ZHOU Yu-long3,WANG Chun-ren3,GONG Da-qing3,QIAN Ai-dong1* (1. College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China,2. Department of Food Engineering,Heilongjiang Land Reclamation Areer College,Harbin 150025,3. College of Animal Science and Veterinary Medcine,Heilongjiang Bayi Agricultural University,Daqing 163319,China) |
| Abstract: | An competitive inhibition ELISA (CI-ELISA) for detection of antibodies against bovine Anaplasma marginale was developed by coating with the recombinant MSP5 protein expressed in E.coli and inhibiting with the monoclonal antibody of 1D8 against the MSP5 protein. The CI-ELISA was specific to inhibit A. marginale positive serum and no crossing inhibitions for sera to Eperythrozoon, Toxophasma, Theileria annulata, and Babesia, respectively. A total of 2,348 bovine serum samples from Heilongjiang area were teste... |
| Keywords: | bovine Anaplasma marginale competitive inhibition ELISA recombinant MSP5 protein epidemiological survey |
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