HCV抗原/抗体联合检测试剂的初步性能评价

Preliminary performance evaluation on a combined HCV-antigen/antibody detection reagent

目的研制HCV抗原/抗体(HCV antigen/antibody, HCV-Ag/Ab)联合检测试剂,并对其性能进行初步评价。方法制备2种HCV多表位嵌合抗原和2株抗HCV核心抗原(HCV core antigen,HCV-cAg)单克隆抗体(单抗),经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳鉴定后,制成HCV双抗原夹心与双抗体夹心ELISA检测试剂。用HCV-Ag/Ab联合试剂对903份血液样品进行检测。对于检测结果与HCV-Ab诊断试剂盒不一致的样品,用MP HCV BLOT 3.0确证试剂确认。采用卡方检验对数据进行分析。结果2种HCV嵌合抗原的相对分子质量约为45 000,与理论预测一致。2株抗HCV-cAg单抗大小正确,且纯度较高。HCV-Ag/Ab联合试剂的特异性达到100%,HCV-Ag检测灵敏度为104 IU/ml,HCV-Ab检测灵敏度为0.1国家临床单位/ml。联合试剂与HCV-Ab诊断试剂盒检测结果的差异无统计学意义(χ^2=0.016 3,P>0.05)。确证试剂对2份HCV-Ab检测不一致样品的确认结果与联合试剂相同。结论制备的HCV-Ag/Ab联合检测试剂特异性强、灵敏度高,适用于血液筛查和临床辅助诊断。

Objective To develop and preliminarily evaluate a combined HCV antigen/antibody (HCV-Ag/Ab) detection reagent.Methods Two HCV multi-epitope chimeric antigens and 2 anti-HCV core antigen (HCV-cAg) monoclonal antibodies were prepared. An HCV double antigen sandwich and double antibody sandwich ELISA reagent was developed after the antigens and antibodies were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The combined HCV-Ag/Ab reagent was used to examine 903 blood samples. The samples of inconsistent results with a HCV-Ab diagnostic kit were validated by MP HCV BLOT 3.0 reagent. The chi-square test was used for data analysis.Results The relative molecular masses of the 2 chimeric antigens were both ~45 000, consistent with the theoretical expectations. Both antibodies had the right size and high purity. The specificity of the combined reagent reached 100%. The sensitivities of HCV-Ag and HCV-Ab detections were 104 IU/ml and 0.1 national clinical unit/ml, respectively. There was no statistical significance between the results of combined reagent and HCV-Ab diagnostic kit (χ^2=0.016 3, P>0.05). The validation reagent confirmed the combined reagent results of 2 samples with discrepant HCV-Ab detection results.Conclusions The combined HCV-Ag/Ab detection reagent has high specificity and sensitivity. It is suitable for blood screening and clinical auxiliary diagnosis.