Development of a quantitative ELISA detection method for Coxsackievirus A group 16 strain(CA16) antigen

目的:建立柯萨奇病毒A组16型(CA16)抗原的双抗体夹心ElISA定量检测方法,用于CA16灭活疫苗的研发和生产过程的抗原定量检测。方法:以CA16中和单抗T26H12为包被抗体、nA14b9为标酶抗体,构建定量检测CA16抗原的双抗体夹心ElISA方法,并对方法的特异性、灵敏度、精密度、准确性、线性和稳定性进行分析。结果:建立了双抗体夹心定量检测CA16抗原的ElISA方法。方法的线性相关系数r2=0.998,线性范围为8~128 ng/Ml,定量限度为8 ng/Ml;变异系数CV80%;与CA16以外的其他样本没有交叉反应。结论:构建的CA16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CA16疫苗的研发和生产过程的抗原活性的定量检测。Objective:To develop an a quantitative enzyme linked immunosorbent assay(Q-ELISA) to determine the concentration of Coxsackievirus A Group 16 Strain(CA16) antigen.This method was used to determine CA16 antigen content at each stage of CA16 vaccine developing and manufacturing process.Methods:A double antibody sandwich Q-ELISA was developed to determine concentration of CA16 antigen,which was based on the high-affinity neutralizing monoclonal antibodies T26H12 as capture antibodies,and NA14B9 as HRP-labeled antibody.The performance of reagent were evaluated.Results:The Q-ELISA for CA16 antigen content was successfully developed.The reagent had good performance.The quantitation scope was 8-128 ng/ml,the coefficient correlation was 0.998,the limit of detection was 8 ng/ml,the recovery was between 87% and 113.8%.The stability was up to 80% after reagent was heated for 6 days at 37℃.The variation coefficient was lower than 15%,and thereagent was no reaction with other sample except CA16 antigen.Conclusion:The Q-ELISA for CA16 antigen was developed with good specificity,accuracy,precision and stability.The method can be used to determine CA16 antigen content during development and production of CA16 vaccine

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JUNO sensitivity on proton decay p → ν K + searches*

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