肠道病毒71型抗原ELISA定量检测方法的建立及应用

发布时间：2018-03-14 12:15

  本文选题：肠道病毒型　切入点：抗原　出处：《微生物学免疫学进展》2016年01期 　论文类型：期刊论文

【摘要】：目的建立肠道病毒71型(EV71)抗原ELISA定量检测方法,用于EV71疫苗生产工艺中抗原定量检测。方法选取抗EV71单克隆抗体作为包被抗体,HRP标记抗EV71多克隆抗体作为酶标二抗,建立EV71抗原ELISA定量检测方法。该方法经系统验证后,应用于定量检测2BS及Vero细胞基质EV71疫苗生产工艺过程样品的抗原含量。结果验证结果显示,该方法的线性范围为3.125~50.000 U/m L,样品回收率为92.0%~110.0%,变异系数小于8.8%;孵育时间及温度在一定范围内偏差的样品回收率为100.8%~113.8%;检测其他肠道病毒中抗原其A值均小于cut-off值;试剂于37℃孵育3 d后的检测样品回收率为101.4%~112.4%;2BS及Vero细胞基质EV71疫苗生产工艺过程样品的适用性检测中,抗原回收率为92.1%~114.9%。结论建立并验证EV71抗原ELISA定量检测方法,其准确度、精密度、耐用性均优良,特异性强、稳定性好,适用于不同细胞基质的EV71疫苗及多价手足口病疫苗生产工艺过程的质量控制。
[Abstract]:Objective to establish a quantitative detection method for ELISA of enterovirus 71 EV71) antigen in the production process of EV71 vaccine. Methods Anti EV71 monoclonal antibody was selected as the anti EV71 polyclonal antibody labeled with anti EV71 polyclonal antibody as enzyme labeled second antibody. A method for quantitative detection of EV71 antigen ELISA was established. The method was used to quantitatively detect the antigen content of 2BS and Vero cell matrix EV71 vaccine samples after systematic verification. The linear range of this method is 3.125 ~ 50.000 UmL, the sample recovery rate is 92.0 and 110.0, the coefficient of variation is less than 8.8, the recovery rate of sample with the deviation of incubation time and temperature is 100.8 ~ 113.8.The A value of antigen in other enterovirus is less than cut-off. After incubated at 37 鈩，

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