摘要
目的 建立一种快速检测肠道病毒71型（EV71） IgM抗体的均相ELISA新技术.方法 首先克隆表达纯化EV71 VP1蛋白,使用生物素标记该蛋白.通过ELISA法摸索出Bio-EV71 VP1的最适抗原量为0.0375 μg.优化供体微球、受体微球、血清等实验反应体系,建立EV71 VP1的均相酶联免疫检测方法的IgM检测技术.使用EV71感染的手足口病（HFMD）患者和健康人血清进行均相酶联免疫检测方法检测评价,并与ELISA检测结果相比较.结果 获得EV71 VP1纯化蛋白并生物素化.摸索出均相酶联免疫检测方法的体系：5 μg/ml受体微球、0.037 5μg生物素化的VP1、20μg/ml供体微球、血清2μl,总体系为20μl.采用ELISA和均相酶联免疫检测方法检测方法对样本血清进行IgM检测.两种方法同时检出手足口病患者lgM阳性16份;14份ELISA法检出阴性样本中,均相酶联免疫检测方法法检出阳性6份.20份健康人血清,2种方法检测结果均为阴性.结论 本研究初步建立了EV71 VP1 IgM的均相酶联免疫检测方法.
Objective To develop new homogeneous enzyme immunosorbent assay for serum IgM antibody of EV71 infection.Methods After protein EV71 VP1 1-297 was purified and biotinylated,the optimal quantities of the antigen was obtained by ELISA.The homogeneous enzyme immunosorbent assay was valuated with the serum samples of both 20 healthy subjects and 30 samples from the patients of HFMD detected by ELISA.Results The reaction system of homogeneous enzyme immunosorbent assay for EV71 IgM was established in this study.Then the homogeneous enzyme immunosorbent assay was valuated with ELISA for EV71 IgM.The results revealed serum IgM were negative for the 20 healthy subjects by both ELISA and homogeneous enzyme immunosorbent assay.Of the 30 HFMD patients,positive serum EV71 IgM detected by both ELISA and homogeneous enzyme immunosorbent assay,and yet,6 IgM positive samples were identified with homogeneous enzyme immunosorbent assay in the 14 negative samples detected by ELISA.Conclusion A homogeneous enzyme immunosorbent assay for detection of serum EV71 IgM has been established.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
2016年第2期220-222,共3页
Chinese Journal of Experimental and Clinical Virology
