抗狂犬病马血清效价ELISA检测试剂的制备  

Preparation of ELISA Kit for Determination of Titer of Horse Antiserum against Rabies Virus

作　　者：薛向光[1] 柳春红[1] 贾媛[1] 张梅[1] 匡科伟[1] 李文娟[1] 郭立君[1] 郭凤云 

机构地区：[1]长春生物制品研究所生化制品室,长春130062

出　　处：《中国生物制品学杂志》2008年第6期519-521,共3页Chinese Journal of Biologicals

摘　　要：目的制备检测抗狂犬病马血清效价的ELISA试剂。方法采用ELISA间接法。先用抗狂犬病病毒IgY包被酶标板,并确定其最适包被浓度;制备酶标二抗,并确定其最佳使用浓度。对试剂的灵敏度、特异性、精密性、稳定性及与小鼠中和试验结果的相关性进行考核。结果抗狂犬病病毒IgY和酶标二抗的最适浓度分别为10!g/ml和1∶1500,试剂灵敏度为0.072IU/ml,特异性良好,批间及试验间变异系数均小于11%,37℃放置3和5d检测样品结果与放置前差异无显著意义,与小鼠中和试验结果呈正相关。结论所制备的抗狂犬病马血清效价ELISA检测试剂盒,可用于抗狂犬病马血清效价的检测。Objective To prepare the ELISA kit for determination of titer of horse antiserum against rabies virus. Methods Optimize the concentration of anti-rabies virus IgY for coating microtiter plate. Prepare enzyme-labeled secondary antibody and optimize its dilution. On the basis of this, an indirect ELISA kit was prepared and evaluated for sensitivity, specificity, precision and stability. Meanwhile, the correlation of determination result by the prepared kit to that by neutralization test was studied. Results The optimal concentration of anti-rabies virus IgY was 10 μg / ml, and the optimal dilution of enzyme-labeled secondary antibody was 1 ： 1 500. The prepared ELISA kit showed good specificity, and its sensitivity was 0. 072 IU/ml. Both inter- and intra-variation coefficients of determination results by the kit were less than 11%. The determination results by the kit after storage at 37℃ for 3 and 5 d showed no significant difference with those before storage. However, the determination result by the kit was positively correlated to that by neutralizing test. Conclusion The prepared ELISA kit may be used for the determination of titer of horse antiserum against rabies virus.

关 键 词：狂犬病病毒 马抗血清 效价 ELISA 

分 类 号：R392-33[医药卫生—免疫学]