百日咳系列抗原的酶联免疫检测法建立及其在组分百日咳疫苗吸附原液中的应用

Establishment of enzyme-linked immunoassay for pertussis series antigen and its application in adsorbed stock solution of component pertussis vaccine

目的:建立百日咳毒素、百日咳丝状血凝素、百日咳黏附素3种抗原含量检测方法,用于组分百日咳疫苗吸附原液的检测。方法:制备抗百日咳毒素、抗百日咳丝状血凝素和抗百日咳黏附素3种兔多克隆抗体,利用棋盘滴定法确定检测条件,建立双抗体夹心酶联免疫吸附法(ELISA法),并进行方法学的验证。结果:3种酶联免疫吸附法的线性相关良好(R^(2)>0.98),线性检测范围分别为1.25~20,2.5~40,0.625~10ng·mL^(-1);该方法的检测准确度高(回收率均在95%~115%),精密度良好(CV<10%)。结论:本研究建立了百日咳系列抗原的酶联免疫吸附法,可用于组分百日咳吸附原液解吸附方法及吸附原液稳定性的研究。

Objective:To established the detection methods for 3 antigens of pertussis toxin,filamentous hemagglutinin and pertactin applied in adsorbed stock solution of pertussis vaccine.Methods:3 rabbit polyclonal antibodies of anti-pertussis toxin,anti-filamentous hemagglutinin and anti-pertactin were prepared.Double-antibody sandwich ELISA was established by checkerboard titration,and the methods were verified.Results:All of the 3 methods had good linear correlation(R^(2)>0.98)within 1.25~20,2.5~40 and 0.625~10 ng·mL^(-1) respectively.The detection accuracies are high(recoveries between 95%and 115%),and the precisions are satisfactory(CV<10%).Conclusion:The enzyme-linked immunoassay for the detection of serial antigens of pertussis were established.It can be used to study the desorption methods and the stability of adsorptive liquid of component pertussis.