IL-6磁微粒化学发光定量检测方法的建立和评价

Development and evaluation of magnetic microparticale chemiluminescence analysis for quantitation of IL-6 in human serum

目的:建立并评价基于磁微粒化学发光免疫分析(CLIA)平台的IL-6定量检测方法。方法:以抗人IL-6抗体包被的磁微粒、待检血清及辣根过氧化物酶标记的IL-6抗体组成分析体系,基于CLIA平台,采用双抗体夹心模式,建立IL-6抗原定量检测方法,并对其进行条件优化和性能评价。结果:该方法的LoB、LoD、FS分别为0.5 pg/ml、2 pg/ml、2.5 pg/ml;分析内、分析间变异及总不精密度分别≤7%、≤10%、≤12%;线性范围为3~1000 pg/ml,最大稀释倍数为8倍;回收率在(100±15)%以内;标本中的血红蛋白、三酰甘油、胆红素浓度分别为200 mg/dl、2000 mg/dl、45 mg/dl时,其检测结果无显著影响;本研究与罗氏同时检测临床标本,其结果的拟合曲线回归方程为:Y=0.955X+9.263,R2=0.9932。结论:本研究建立的IL-6磁微粒化学发光定量检测方法性能良好,具有一定的临床意义,可满足临床检测IL-6水平的需要。

Objective:To establish and evaluate quantitative detection method for IL-6 based on magnetic microparticale chemilumineseent immunoassay(CLIA).Methods:The analysis system was composed of magnetic microparticale coated with anti-human IL-6 antibody,serum and anti-human IL-6 antibody labeled with horseradish peroxidase.Based on the CLIA platform,the double antibody sandwich mode was adopted to test quantitatively of IL-6 antigen.Moreover,the reaction conditions of IL-6 were optimized and the analytical performance was evaluated.Results:It was indicated that the limit of blank(LoB),limit of detection(LoD)and functional sensitivity(FS)were 0.5,2 and 2.5 pg/ml.The precision of the within variantion and interanalytical variantion and total imprecision were lower than 7%,10%,12%,respectively.The linear range was within the range between 3 and 1000 pg/ml,and the maximum dilution limit was 1∶7.The recovery was within(100±15)%.The results measured by adding lower than 200 mg/dl hemoglobin,2000 mg/dl triglyceride and 45 mg/dl bilirubin to human sera with high and low concentrations of IL-6 were undisturbed.The clinical serum samples were tested simultaneously by this method and Roche system,and the curve regression equation of the results was Y=0.955 X+9.263,R2=0.9932.Conclusion:The quantitative detection method for IL-6 introduced in this study was demonstrated to have effective performance and can meet the need of clinical requirement.