Cox A16抗原检测试剂研究及初步应用  

Research and preliminary application of Cox A16 antigen detection kit

作　　者：刘正玲[1] 李华[1] 龙润乡[1] 杨婷[1] 杨蓉[1] 岳磊[1] 谢忠平[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所,云南省重大传染病疫苗工程技术研究中心,云南省重大传染病疫苗研发重点实验室,昆明650118

出　　处：《中国卫生检验杂志》2013年第4期801-805,共5页Chinese Journal of Health Laboratory Technology

基　　金：云南省重点新产品研究项目(2012BC006)

摘　　要：目的:建立柯萨奇病毒A组16型抗原检测酶联免疫法,为疫苗研发及病原学检查提供有效的技术手段。方法:以CA16多抗为包被抗体,以辣根过氧化物酶标记抗体为指示物建立CA16抗原ELISA检测法,对其线性、灵敏度、特异度、精密度及稳定性等指标进行分析。将所建立的方法应用到疫苗研发及病原检测中进行验证及评价。结果:建立的CA16抗原ELISA检测线性范围为1.25μg/ml～80μg/ml,R2达0.98以上;最低检测下限为1.25μg/ml,可检测到2×102CCID50～9×103CCID50活病毒;与CA16以外的10种肠道病毒无交叉反应;变异系数CV小于15%。与荧光PCR、中和法鉴定结果一致性高。结论:建立的ELISA法各项性能符合抗原检测要求,可用于CA16疫苗研发及病原学检查。Objective：To develop an Enzyme Linked Immuno-sorbent Assay（ELISA） for CA16 vaccine research and CA16 antigen detection.Methods： A double antibody sandwich ELISA method was developed by using polyclonal antibody as coating antibody and HRP labeled antibody as enzyme labeled antibody.The method was then verified,applied and evaluated.Results： The linear range of the developed ELISA method was 1.25 μg/ml^80 μg/ml,R2 value was 0.98＋.The quantitation limit of the developed method was 1.25 μg/ml,and the detection limit of live CA16 in tissue culture was about 2×102 CCID50~9×103 CCID50.The method showed no cross reactions with the other ten kinds of enterovirus except CA16.The variation coefficient was lower than 15%.The test results was higher consistency with that of fluorescence PCR and neutralization test.Conclusion： The developed ELISA showed good linearity,sensitivity,specificity,precision and stability.The method can be used to develop CA16 vaccine and to detect CA16 antigen.

关 键 词：手足口病 肠道病毒 柯萨奇病毒A组16型 

分 类 号：R446.5[医药卫生—诊断学]