目的  建立并部分验证水痘-带状疱疹病毒糖蛋白E（glycoprotein E, gE）ELISA。方法  用中国仓鼠卵巢细胞表达体系表达水痘-带状疱疹病毒的gE抗原，3步层析纯化后，混合弗氏佐剂免疫BALB/c小鼠和日本大耳白兔，制备抗gE特异性多克隆抗体。制备的抗体经Protein A亲和层析纯化后，用于建立双抗体夹心ELISA检测gE抗原。考察方法的线性范围、特异性、准确性、重复性和中间精密度等。结果  表达的gE经3步层析纯化后，纯度达到99%以上。该方法在gE浓度2^﹣2~2⁵ μg/ml范围内线性关系良好，决定系数大于0.99；该方法可特异性识别gE，与中国仓鼠卵巢细胞上清液或牛血清白蛋白溶液无交叉反应；高、中、低浓度gE内部参考品检测均值回收率分别为99.44%、94.35%、94.37%，变异系数分别为6.07%、4.28%、12.62%；重复性验证变异系数为8.24%，中间精密度验证变异系数为7.55%。结论  建立的双抗体夹心ELISA可以用于水痘-带状疱疹病毒gE的定量检测及疫苗质量标准的建立。

Objective  To establish and partially validate ELISA for varicella-zoster virus (VZV) glycoprotein E (gE) detection. Methods  The gE antigen of VZV was expressed by the Chinese hamster ovary (CHO) cell expression system. After three-step chromatography purification, gE was mixed with Freund's adjuvant to immunize BALB/c mice and Japanese white rabbits for the preparation of gE specific polyclonal antibodies. The prepared antibodies were purified by Protein A chromatography and used to establish the double-antibody sandwich ELISA for gE antigens detection. The linear range, specificity, accuracy, repeatability, and intermediate precision of the method were verified.  Results  After three-step chromatography purification, the purity of gE antigen reached more than 99%. The method showed good linearity in gE concentration range of 2^﹣2-2⁵ μg/ml, and coefficient of determination was >0.99. This method specifically detected gE with no cross reaction with CHO cell supernatant or bovine serum albumin solution. The mean recovery rates of gE internal reference with high, medium, and low concentrations were 99.44%, 94.35%, 94.37%, and coefficients of variation (CVs) were 6.07%, 4.28%, 12.62%, respectively. The repeatability verification CV was 8.24%, and the intermediate precision verification CV was 7.55%. Conclusion  The established double-antibody sandwich ELISA can be used for the quantitative detection of VZV gE antigen and the establishment of vaccine quality control standards.