| 蓝舌病病毒4型特异性竞争ELISA检测方法的建立 |
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| 引用本文： | 李佳璇,臧明鑫,谢双羽,姜艳平,崔文,徐义刚,刘敏,乔薪瑗,王丽,周晗,李一经,唐丽杰.蓝舌病病毒4型特异性竞争ELISA检测方法的建立[J].生物工程学报,2017,33(8):1284-1291. |
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| 作者姓名： | 李佳璇 臧明鑫 谢双羽 姜艳平 崔文 徐义刚 刘敏 乔薪瑗 王丽 周晗 李一经 唐丽杰 |
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| 作者单位： | 东北农业大学 动物医学学院，黑龙江 哈尔滨 150030,东北农业大学 动物医学学院，黑龙江 哈尔滨 150030,东北农业大学 动物医学学院，黑龙江 哈尔滨 150030,东北农业大学 动物医学学院，黑龙江 哈尔滨 150030,东北农业大学 动物医学学院，黑龙江 哈尔滨 150030,东北农业大学 动物医学学院，黑龙江 哈尔滨 150030,东北农业大学 动物医学学院，黑龙江 哈尔滨 150030,东北农业大学 动物医学学院，黑龙江 哈尔滨 150030,东北农业大学 动物医学学院，黑龙江 哈尔滨 150030,东北农业大学 动物医学学院，黑龙江 哈尔滨 150030,东北农业大学 动物医学学院，黑龙江 哈尔滨 150030,东北农业大学 动物医学学院，黑龙江 哈尔滨 150030 |
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| 基金项目： | 国家科技支撑计划 (No. 2013BAD12B01) 资助。 |
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| 摘 要： | 旨在建立蓝舌病病毒4型(BTV-4)特异性抗体ELISA检测方法,为蓝舌病的免疫学诊断提供新的技术。利用制备的两株抗4型BTV VP2蛋白的单克隆抗体4A-1G7和4B-1B6,建立BTV-4特异性竞争ELISA抗体检测方法。利用该方法同时对50份羊和牛BTV阴性血清进行检测,分别确定两种方法阻断率临界值为49%和40%。利用标准阳性血清检测的试验结果表明,该方法的敏感性、特异性和重复性符合OIE通用标准。同时,4A-1G7和4B-1B6两种竞争ELISA方法联合作用,可以检测感染4、18和20型BTV的血清。研究结果为建立以上各型BTV的检测方法提供了基础。
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| 关 键 词： | 蓝舌病病毒，竞争ELISA，单克隆抗体，VP2蛋白 |
| 收稿时间： | 2017/3/25 0:00:00 |
Establishment of two competitive ELISAs for specific detection of bluetongue virus serotype 4 |
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| Jiaxuan Li,Mingxin Zang,Shuangyu Xie,Yanping Jiang,Wen Cui,Yigang Xu,Min Liu,Xinyuan Qiao,Li Wang,Han Zhou,Yijing Li and Lijie Tang.Establishment of two competitive ELISAs for specific detection of bluetongue virus serotype 4[J].Chinese Journal of Biotechnology,2017,33(8):1284-1291. |
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| Authors: | Jiaxuan Li Mingxin Zang Shuangyu Xie Yanping Jiang Wen Cui Yigang Xu Min Liu Xinyuan Qiao Li Wang Han Zhou Yijing Li and Lijie Tang |
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| Institution: | College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, Heilongjiang, China,College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, Heilongjiang, China,College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, Heilongjiang, China,College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, Heilongjiang, China,College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, Heilongjiang, China,College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, Heilongjiang, China,College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, Heilongjiang, China,College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, Heilongjiang, China,College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, Heilongjiang, China,College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, Heilongjiang, China,College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, Heilongjiang, China and College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, Heilongjiang, China |
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| Abstract: | To develop a clinical diagnosis technique for bluetongue virus infection, we established serotype-specific methods to detect serotype 4 of bluetongue virus (BTV-4). Two monoclonal antibodies (mAbs) against VP2 protein of BTV-4, named 4A-1G7 and 4B-1B6, were used as competitive antibodies in the competitive enzyme-linked immunosorbent assays (C-ELISA). We detected 50 negative serum samples from sheep, goats and cattle by C-ELISA. The cut-off values of 4A-1G7 and 4B-1B6 mAbs were 49% and 40%, respectively. The results of the sensitivity, specificity and repeatability by detecting standard positive serum, were consistent with the general standard of Office International Des Epizooties. Furthermore, serum samples of BTV-4, BTV-18 and BTV-20 infection could be screened out through the combined C-ELISAs by 4A-1G7 and 4B-1B6 mAbs. Thus, this technique may diagnose BTV-4, BTV-18 and BTV-20 infections. |
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| Keywords: | bluetongue virus serotype 4 competitive ELISA monoclonal antibody VP2 protein |
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