肠道病毒71型抗原含量检测方法的建立及初步验证

Development A method of quantitative assay for enterovirus 71

目的建立肠道病毒71型（EV71）抗原含量检测方法并进行初步验证，用于疫苗研制及生产过程中抗原含量的检测。方法以抗EV71单克隆抗体为包被抗体，酶标抗EV71多克隆抗体为检测抗体，建立双抗体夹心ELISA方法，以本室自制EV71抗原参考品为定量标准，建立抗原标准曲线，确定定量限度及最佳定量范围，并对该方法的准确度、精密度和特异性进行初步验证。结果建立了双抗体夹心检测EV71抗原含量的ELISA方法，定量限度为0．23μg／ml，最佳定量范围为7．32～0．23μg／ml，相关系数为R^2=0．9976；准确性较好，回收率在90％～110％之间；精密性较好，变异系数小于10％；除与EV71样品有特异性反应外，与其他样品无交叉反应。结论建立了检测EV71抗原含量的双抗体夹心ELISA方法并进行了初步验证，为疫苗研制过程中抗原含量测定提供了简便、快捷的检测手段。

Objective To establish a quantitative assay for enterovirus 71, this can be used in detecting the virus content during vaccine development and production. Methods We established the method of qaantitafive assay for EV71 by using double antibody sandwich ELISA. The sensitivity, accuracy, precision and specificity of the method were evaluated. Results We developed an ELISA method to quantitative assay for EVT1. The quantitation limit of the method is 0.23 μg/ml and the quantitation scope of the method is 7.32--0.23 μg/ml, the coefficient correlation is R2 = 0.9976;The method showed good accuracy, precision and specificity. The recovery is between 90%--110% and the variation coefficient is lower than 10%. Conclusion An ELISA method was developed for the quantitative assay of EV71 virus, which can be used for the rapid quantitative determination of EV71 virus during vaccine development and production.