狂犬病病毒抗原ELISA检测方法的建立及其应用  

Development and Application of ELISA Method for Rabies Virus Antigen

作　　者：王亚军[1,2] 戚凤春[1] 薛向光[1] 李晓波[1] 王丽娜[1] 谢琳[1] 王宇[1] 夏青娟[1] 隋波 张梅[1] 魏涛 郭立君[1] 

机构地区：[1]长春生物制品研究所,长春130062 [2]吉林大学生命科学学院,长春130023 [3]长春祈健生物有限公司,长春130000 [4]长春长生基因药业股份有限公司,长春130000

出　　处：《中国生物制品学杂志》2009年第7期713-715,共3页Chinese Journal of Biologicals

摘　　要：目的建立狂犬病病毒抗原ELISA检测方法,并进行初步应用。方法采用狂犬病病毒CTN-1V纯化抗原免疫家兔,制备多克隆抗体,作为酶标抗体,马抗狂犬血清纯化抗体作为包被抗体,建立双抗体夹心ELISA法。对该方法进行验证,检测狂犬病病毒原液毒力和疫苗效力,并与小鼠(NIH)法检测结果进行对比。结果双抗体夹心ELISA检测方法的最低检出限为0.17 IU/ml,最佳线性范围为0.17～2.00 IU/ml,相关系数R>0.97;批间及试验内变异系数均小于10%,特异性、稳定性均良好;检测20批病毒原液毒力及疫苗效力,结果与NIH法相比,差异无统计学意义。结论已建立狂犬病病毒抗原ELISA检测方法,可用于生产过程中狂犬病疫苗原液和半成品的检定。Objective To develop and preliminarily apply an ELISA method for rabies virus antigen. Methods Polyclonal antibody was prepared by immunizing rabbits with purified rabies virus CTN-1V antigen. A double antibody sandwich ELISA method was developed by using the prepared polyclonal antibody as labeling antibody and purified horse antisera against rabies as coating antibody, then verified and used for determination of virulence of virus bulk and potency of vaccine. The determination result was compared with that by NIH test. Results The minimum detection limit, optimal linear range and correlation coefficient of the developed ELISA method were 0. 17 IU/ml, 0. 17 - 2. 00 IU/ml and more than 0. 97 respectively, and both inter- and intra-variation coefficients were less than 10%. The developed ELISA method showed good specificity and stability. The determination results of 20 batches of virus bulks and prepared vaccines by the developed method showed no significant differences with those by NIH method. Conclusion An ELISA method for rabies virus antigen is developed, which may be used for the control tests on bulk and final bulk during production of rabies vaccine.

关 键 词：狂犬病疫苗 病毒抗原 ELISA NIH法 效力 

分 类 号：R373.9[医药卫生—病原生物学] R392.33[医药卫生—基础医学]