| 泰勒焦虫重组蛋白 TaSP-Tams1-SPAG1间接ELISA 检测方法的建立 |
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| 引用本文： | 魏玉荣,易忠,马文戈,郭会玲,向银珍,米晓云,陈智,魏婕,苗书魁,吴国梁,祁巧芬,埃尼瓦尔·太外库力,黄炯. 泰勒焦虫重组蛋白 TaSP-Tams1-SPAG1间接ELISA 检测方法的建立[J]. 新疆农业大学学报, 2013, 0(5): 360-365 |
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| 作者姓名： | 魏玉荣 易忠 马文戈 郭会玲 向银珍 米晓云 陈智 魏婕 苗书魁 吴国梁 祁巧芬 埃尼瓦尔·太外库力 黄炯 |
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| 作者单位： | [1]新疆畜牧科学院兽医研究所,乌鲁木齐830000 [2]吐鲁番地区动物疾病控制与诊断中心,吐鲁番838000 [3]新疆乌鲁木齐县动物卫生监督所,乌鲁木齐830037 [4]新疆喀什地区动物疾病控制与诊断中心,喀什844000 |
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| 基金项目： | 新疆维吾尔自治区自然基金青年基金项目（2011211B38）,国家自然科学基金项目（31160505） |
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| 摘 要： | 以真核表达的焦虫表面抗原重组蛋白 TaSP-Tams1-SPAG1作为 ELISA 板包被抗原,通过方阵实验确定抗原包被浓度,探索抗原包被时间、温度、一抗及二抗血清稀释度及作用时间,建立牛环形泰勒焦虫表面抗原重组蛋白 ELISA 方法.结果得出 TaSP-Tams1-SPAG1的最佳包被浓度为10μg/mL,血清最佳稀释度为1∶100, ELISA 阳性反应的临界值为 OD 492nm ≥0．348,重复试验变异系数小于12％,所建立的 ELISA 方法不与牛巴贝斯虫及牛瑟氏泰勒虫血清发生反应.TaSP-Tams1-SPAG1为抗原建立的 ELISA 方法特异性强,敏感度高,重复性好,为牛环形泰勒焦虫病的准确检测及防治提供技术支持.
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| 关 键 词： | 泰勒焦虫 TaSP-Tams1-SPAG1 重组蛋白 ELISA |
Development of ELISA with Recombinant Protein Antigens for Theileria annulata |
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| WEI Yu-rong,YI Zhong,MA Wen-ge,GUO Hui-ling,XIANG Yin-zhen,MI Xiao-yun,CHEN Zhi,WEI Jie,MIAO Shu-kui,WU Guo-liang,QI Qiao-fen,Ainiwaer Taiwaikuli,HUANG Jiong. Development of ELISA with Recombinant Protein Antigens for Theileria annulata[J]. Journal of Xinjiang Agricultural University, 2013, 0(5): 360-365 |
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| Authors: | WEI Yu-rong YI Zhong MA Wen-ge GUO Hui-ling XIANG Yin-zhen MI Xiao-yun CHEN Zhi WEI Jie MIAO Shu-kui WU Guo-liang QI Qiao-fen Ainiwaer Taiwaikuli HUANG Jiong |
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| Affiliation: | WEI Yu-rong,YI Zhong,MA Wen-ge,GUO Hui-ling,XIANG Yin-zhen,MI Xiao-yun,CHEN Zhi,WEI Jie,MIAO Shu-kui,WU Guo-liang,QI Qiao-fen,Ainiwaer Taiwaikuli,HUANG Jiong |
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| Abstract: | The objective of this paper was to discuss the antigen coating time,temperature,serum dilution of first antibody and second antibody and action time,and develop antigen recombinant protein ELISA method against Theileria annulata in cattle,to determine antigen coating concentration on the square ex-periment by using antigen recombinant protein TaSP-Tams1-SPAG1 against Theileria annulata expressed by eukarya as ELISA coating antigen.The results indicated that the optimal concentration of coating anti-gen was 10 μg/mL and the optimal blocking dilution of serum sample was 1 ∶100 in the cross assay.The critical value reacted by ELISA positive was an OD 492nm ≥ 0.348.The variation coefficient of intra-batch and the inter-batch in the repeating tests was less than 12%.No cross reactions were found among the piro-plasms.The improved indirect TaSP-Tams1-SPAG1 ELISA Was highly sensitive specific and reproducible, which provided a technical basis for accurate detection and control of Theileria annulata diseases in cattle. |
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| Keywords: | Theileria annulata TaSP-Tams1-SPAG1 recombination protein ELISA |
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