百日咳毒素S1亚基蛋白时间分辨免疫荧光分析法的建立及应用

Development of a time-resolved fluoroimmunoassay for detecting S1 subunit of pertussis toxin and its application

目的建立铕(Eu3+)标记检测百日咳毒素S1亚基蛋白时间分辨免疫荧光分析法。方法利用原核重组表达系统表达百日咳毒素S1亚基蛋白,并鉴定,采用双抗体夹心法建立检测S1 TRFIA分析方法。结果自制S1试剂盒检测灵敏度2.5 ng/ml,与其他抗原无交叉反应,并初步可应用于百日咳疫苗中免疫原的监测。结论自制百日咳毒素S1亚基蛋白时间分辨免疫荧光分析法能够较为可靠的监控百日咳疫苗质量。

Objective To develop a time-resolved fluoroimmunoassay（TRFIA） for detection of pertussis toxin（PT） S1 subunit for quality control of human PT vaccine.Methods A double antibody sandwich one-step method was used to establish the TRFIA for detecting PT S1 subunit in the vaccine.Results The sensitivity of c peptide analysis reached 2.5 ng/ml without cross-reactions with other antigens.This assay could be used in detecting S1 subunit in the vaccine.Conclusion The TRFIA for detecting PT S1 subunit is simple,sensitive and rapid for quality control of the PT vacccine.