快速化学发光高敏C-反应蛋白定量试剂盒的研制

Development of a rapid chemiluminesent immunoassay test kit for quantitative determination of high sensitivity C-reactive protein

目的快速化学发光高敏C-反应蛋白定量试剂盒的研制及评价。方法建立以链霉亲和素包被微孔板,生物素连接一株单克隆抗体,辣根过氧化物酶标记另一株单克隆抗体,鲁米诺和过氧化脲为发光底物的化学发光定量试剂盒,对反应条件进行优化,并对性能参数作出评价。结果该试剂盒敏感度为0.005mg/L,线性范围为1-500mg/L,批内变异系数为5.4%-6.4%,批间变异系数为6.0%-7.8%,相对偏倚小于4.2%,165例临床标本的测试结果显示该试剂与国外试剂的测定结果Youden指数为0.951 1,Kappa指数为0.951 5。结论该试剂盒灵敏度高,有良好的准确性和重复性,线性范围宽,符合临床检测需要。

Objective To develop and evaluate a rapid chemiluminesent immunoassay（CLIA）test kit for quantitative determination of high sensitivity C-reactive protein in human serum.Methods Coated microplate with streptavidin,the double antibody sandwich method was used to develop the high sensitivity C-reactive protein kit,biotinylated one monoclonal antibody and labeled the other monoclonal antibody with horseradish peroxidase,luminol and peroxide was used for substrate system,optimized the reaction condition,Then the methodological evaluation was assessed.Results The sensitivity was reached to 0.005mg/L,The linear range of the kit was 1-500mg/L,the intra and inter-assay coefficient of variation were 5.4%-6.4% and 6.0%-7.8% respectively,The reagents kit of bias was less than 4.2%.The Youden index between the developmental assay and imported kit for high sensitivity C-reactive protein of 165blood samples was 0.951 1,Kappa index was 0.951 5.Conclusion The developed CLIA kit for the detection of high sensitivity C-reactive protein in human serum is a sensitive method with broad linear range,high accuracy and reproducibility.It could meet the clinical requirements for the detection of the concentration.