一种新的高纯度蓝舌病毒中国湖北株制备方法  

Purification of Bluetongue Virus Strain HbC by a Novel Co-immunoprecipitaion Method

作　　者：卢莉莉[1] 董长垣[1] 郭淑芳[1] 罗翔[1] 张蔚英[1] 刘文培[1] 

机构地区：[1]武汉大学医学院病毒学研究所分子病毒与癌研究室,武汉430071

出　　处：《武汉大学学报（医学版）》2004年第3期335-338,共4页Medical Journal of Wuhan University

基　　金：湖北省教育厅重点项目 ;批号 :2 0 0 0B0 3 0 10

摘　　要：目的 :获得高纯度蓝舌病毒HbC株 ,为制备生产和实验室研究奠定基础。方法 :利用反向免疫共沉淀方法获取纯化的蓝舌病毒颗粒 ;运用透射电镜和凝胶过滤层析检测纯化产物病毒的结构完整性和纯度 ;组织培养技术检测产物的生物学活性并计算纯化得率。结果 :纯化后的病毒经凝胶过滤层析分析仅见一个层析峰 ;所对应的电镜照片背景干净 ,未见明显杂质 ;纯化后的病毒对Vero细胞的TCID50 是 1 0 -5.7·ml-1 ;纯化得率为 5 0 .4 %～70 .8%。结论 :这种新方法能制备出高纯度、高活性的蓝舌病毒 ,且得率较高 ,易于放大生产 。Objective: To get the high purified bluetongue virus strain HbC for further study and product.Methods: A novel reverse co immunoprecipitation method was used to get the purified BTV HbC particles and gel filtration, electron microscopy(EM), TCID 50 and plaque assay was used to analyse the purification results.Results: The gel filtration analysis of purified virus showed only one absorbance peak; the background of it’s electron micrograph was clean; TCID 50 of purified virus on Vero cells was 10 -5.7 ·ml -1 ; and the recovery rate was within 50.4%～70.8%.Conclusion: The new method introduced can produce large amounts of high purified viral particles with high infectivity so it can be applied to further reseach on developing a large scale production system.

关 键 词：蓝舌病毒中国湖北株 反向免疫共沉淀法 溶癌病毒 凝胶过滤层析 

分 类 号：Q939.44[生物学—微生物学]