HCV核心抗原磁微粒化学发光法检测试剂的研制及初步性能评价

Development and preliminary performance evaluation of HCV core antigen chemiluminescence microparticle immunoassay detection reagent

目的研制丙型肝炎病毒核心抗原(HCV core antigen,HCV-cAg)磁微粒化学发光法(chemiluminescence microparticle immunoassay,CMIA)的检测试剂,并初步评价其性能。方法采用2株抗HCV-cAg单克隆抗体(单抗)预先包被磁微粒,另2株抗HCV-cAg单抗用吖啶酯标记,制备双抗体夹心CMIA检测试剂;与ELISA试剂盒同时检测210例干扰物样本、1158例健康人样本、140例HCV-RNA阳性样本和2套血清转换盘样本,鉴定研制试剂的特异性和灵敏度。结果CMIA检测试剂的特异性为99.74%,ELISA试剂盒的特异性为99.57%,2种方法干扰物样本的阴性率均为100%;CMIA检测试剂的灵敏度为77.86%,ELISA试剂盒的灵敏度为62.14%,且CMIA检测试剂检出血清转换盘样本阳性的时间早于ELISA试剂盒。结论成功制备了HCV-cAg CMIA检测试剂,该试剂的特异性好、灵敏度高,适用于HCV的早期筛查和临床辅助诊断。

Objective To develop a chemiluminescence microparticle immunoassay(CMIA)detection reagent for hepatitis C virus core antigen(HCV-cAg)and preliminarily evaluate its performance.Methods Two anti-HCV-cAg monoclonal antibodies(Mabs)were pre-coated with magnetic particles,and the other two anti-HCV-cAg Mabs were labeled with acridine ester to prepare a CMIA detection reagent for double-antibody sandwich magnetic particles.Using CMIA and ELISA kit,210 interference samples,1158 normal human samples,140 HCV-RNA positive samples,and 2 sets of serum conversion disk samples were detected simultaneously to identify the specificity and sensitivity of the developed reagent.Results The specificity of CMIA detection reagent was 99.74%,and the specificity of ELISA kit was 99.57%.The negative rates of both methods for detecting interference samples were 100%.The sensitivity of CMIA detection reagent was 77.86%,the sensitivity of ELISA kit was 62.14%,and the CMIA detection reagent detected positive serum conversion disk samples earlier than ELISA kit.Conclusion HCV-cAg CMIA detection reagent is successfully prepared with good specificity and high sensitivity,and is suitable for early screening and clinical auxiliary diagnosis of HCV.