| 牛环形泰勒虫GST-Tams1融合蛋白间接ELISA检测方法的建立 |
| |
| 引用本文： | 简子健,马素贞,黄家雨,孙其喆,沈炯玉,苗中秋,吕伟.牛环形泰勒虫GST-Tams1融合蛋白间接ELISA检测方法的建立[J].新疆农业科学,2011,48(3):578-583. |
| |
| 作者姓名： | 简子健 马素贞 黄家雨 孙其喆 沈炯玉 苗中秋 吕伟 |
| |
| 作者单位： | 新疆农业大学动物医学学院,乌鲁木齐,830052 |
| |
| 摘 要： | 目的与方法]以纯化的牛环形泰勒虫GST-Tams1融合蛋白作为检测抗原,通过优化ELISA反应条件,建立检测牛环形泰勒虫血清特异性抗体的间接ELISA方法.结果]方阵试验确定的GST-Tams1抗原的最适包被浓度为10 μg/mL,血清最佳稀释倍数为80倍,ELISA阳性反应的临界值为OD<,450>≥0.282,批内和批间重复试验的变异系数均小于15;.Tams1间接ELISA方法能排除GST的干扰,与其它梨形虫病无交叉反应,与环形泰勒虫病巢式PCR检测方法的阳性符合率为96;.结论]建立的Tams1间接ELSIA检测法重复性好、特异性强、灵敏度高.这是国内首次利用重组蛋白建立的牛环形泰勒虫病血清学诊断方法,为大规模地进行牛环形泰勒虫病的流行病学调查和血清学诊断提供有效的技术手段.
|
| 关 键 词： | 牛环形泰勒虫 裂殖子表面抗原1 重组抗原间接ELISA 建立 |
| 收稿时间： | 2011-03-25 |
Development of an Improved Indirect Tams1 ELISA for Detection of Theileria Annulata in Cattle |
| |
| JIAN Zi-jian,MA Su-zhen,HUANG Jia-yu,SUN Qi-zhe,SHEN Jiong-Yu,MIAO Zhong-qiu,L Wei.Development of an Improved Indirect Tams1 ELISA for Detection of Theileria Annulata in Cattle[J].Xinjiang Agricultural Sciences,2011,48(3):578-583. |
| |
| Authors: | JIAN Zi-jian MA Su-zhen HUANG Jia-yu SUN Qi-zhe SHEN Jiong-Yu MIAO Zhong-qiu L Wei |
| |
| Institution: | JIAN Zi-jian,MA Su-zhen,HUANG Jia-yu,SUN Qi-zhe,SHEN Jiong-Yu,MIAO Zhong-qiu,L(U) Wei |
| |
| Abstract: | 【Objective and Method】An improved indirect Tams1 enzyme-linked immunosorbent ELISA assay(indirect Tams1 ELISA) was developed for the detection of specific antibody against Theileria annulata in cattle after optimizing its reacting conditions.【Result】The optimal concentration of coating antigen was 10 μg/mLand the optimal dilution of serum sample was 1:80 in the cross assay.The cutoff was chosen as an OD450≥0.282 for positive response.The variation coefficient of intra-batch and the inter-batch in the repeating tests was less than 15%.The interference of murine GST antiserum could be eliminated and no cross reactions were found among the piroplasms.The coincidences of identified positive serum samples for bovine piroplasmosis in indirect Tams 1 ELISAs were 96%,comparing with that in nested PCR.【Conclusion】The improved indirect Tams1 ELISA was highly sensitive,specific and reproducible.This was the first case to establish an improved indirect Tams1 ELISA for detection of specific antibodies against Theileria annulata in China,which provided a new tool for the large-scale epidemiological surveys and serological diagnosis of bovine theileriosis infected with Theileria annulata. |
| |
| Keywords: | Theileria annulata Tams1 recombination protein indirect Tams1 ELISA development |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《新疆农业科学》浏览原始摘要信息 |
| 点击此处可从《新疆农业科学》下载免费的PDF全文 |
|
