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目的:建立水痘-带状疱疹病毒(VZV)抗原的双抗体夹心ELISA定量检测方法,用于质控VZV灭活疫苗研发和生产中抗原含量。方法:以VZV中和单抗5F6C8为包被抗体、8H5D1为酶标抗体,构建定量检测VZV抗原的双抗体夹心ELISA方法,并对本方法的特异性、灵敏度、准确性、线性和稳定性等性能进行分析。结果:建立的双抗体夹心定量检测VZV抗原的ELISA方法,线性范围为0.4μg～13μg/ml,相关系数为R2=0.994,定量限度为0.4μg/ml;变异系数CV<15%、准确性回收率介于87.5%～111.6%之间,稳定性37℃6天的回收率>80%。与VZV以外的相关病毒样本没有交叉反应。结论:构建的VZV抗原ELISA定量检测方法的各项性能符合定量检测需要,可用于VZV灭活疫苗的研发和生产过程的抗原含量检测。
OBJECTIVE: To establish a double-antibody sandwich ELISA for the quantitative determination of varicella-zoster virus (VZV) antigen and to control the antigen content in the development and production of VZV inactivated vaccine. Methods: VZV neutralizing monoclonal antibody 5F6C8 was used as coating antibody and 8H5D1 as enzyme-labeled antibody to construct double-antibody sandwich ELISA for the quantitative detection of VZV antigen. The specificity, sensitivity, accuracy, linearity and stability of this method Analyze. Results: The established ELISA method for the quantitative detection of VZV antigen by sandwich ELISA was linear in the range of 0.4μg ~ 13μg / ml, with a correlation coefficient of R2 = 0.994 and a limit of quantification of 0.4μg / ml. The coefficient of variation CV <15% Rates ranged from 87.5% to 111.6%, with a recovery of> 80% at 37 ° C for 6 days. No cross-reactivity with relevant virus samples other than VZV. Conclusion: The performance of quantitative VZV antigen ELISA assay meets the needs of quantitative detection. It can be used to detect the antigen content of VZV inactivated vaccine.