基于吖啶酯化学发光免疫定量分析技术建立血清IL-6水平检测方法与初步应用评价  

Establishment of the Detection Method of Serum IL-6 Level and Its Preliminary Application Evaluation based on Acridine Ester Chemiluminescence Immune Quantitative Analysis Technology

作　　者：唐红辉 李洪春 TANG Honghui;LI Hongchun(College of Medical Technology,Xuzhou Medical University,Jiangsu Xuzhou 221000,China;Clinical Testing Center,the Fourth Affiliated Hospital of Soochow University/Suzhou Dushu Lake Hospital,Jiangsu Suzhou 215000,China)

机构地区：[1]徐州医科大学医学技术学院,江苏徐州221000 [2]苏州大学附属第四医院、苏州市独墅湖医院临床检测中心,江苏苏州215000

出　　处：《现代检验医学杂志》2024年第4期175-179,185,共6页Journal of Modern Laboratory Medicine

基　　金：苏州市科技计划项目(SZM2021011)。

摘　　要：目的基于吖啶酯化学发光免疫定量分析技术,建立一种血清白细胞介素-6(interleukin 6,IL-6)的检测方法。方法采用双抗体夹心法,分别用吖啶酯标记完整抗体和生物素标记酶切片段抗体,并与待测物IL-6形成夹心复合物,其中生物素再与链霉亲和素包被的磁性固相微粒特异反应发光,通过测量发光信号值进行定量分析;并对其标记抗体稀释缓冲液、标记抗体浓度、样本加样量等进行条件优化;对空白限(LOB)、检出限(LOD)、中间精密度、可报告范围、干扰实验、钩状(HOOK)效应等性能指标进行评价;选择145例血清样本,采用线性回归分析与罗氏电化学发光法进行方法学比对。结果成功建立一种基于吖啶酯化学发光免疫定量分析技术检测血清IL-6的方法。该方法选择MESBSA作为标记抗体稀释缓冲液,标记抗体浓度选择0.2μg/ml,样本加样量选择50μl。该方法的LOB为0.2 pg/ml,LOD为0.5 pg/ml,可报告范围为0.5~30000 pg/ml,中间精密度<5.3%。在IL-6达到200000 pg/ml浓度水平时,未出现HOOK效应现象。在三酰甘油(TG)30000μg/ml,血红蛋白(HGB)9000μg/ml,胆红素(TBIL)330μg/ml,类风湿因子(RF)1500 IU/ml浓度范围内,干扰物质对检测结果无影响。与电化学发光法测定IL-6比对,线性回归方程为Y=0.9802X-3.4879,r=0.9977,两种方法试剂测定结果呈高度相关(P<0.05)。结论建立了基于吖啶酯化学发光检测血清IL-6的方法,各项指标均符合临床应用要求,适宜在临床实验室推广应用。Objective To establish a method for the detection of serum interleukin(IL)-6 based on acridine ester chemiluminescence immune quantitative analysis.Methods Double-antibody sandwich method was applied,acridine ester was used to label complete antibody,and biotin was used to label enzyme-cut fragment antibody.Next,then they formed a sandwich complex with tested substance IL-6,in which biotin and streptavidin-coated magnetic solid particles reacted specifically to luminescence.Quantitative analysis was conducted by measuring the luminescence signal value,and the conditions for labeling antibody dilution buffer,concentration of labeled antibody and sample addition amount were optimized.The performance indexes such as limit of blank(LOB),limit of detection(LOD),intermediate precision,reportable range,interference test and HOOK effect were evaluated.Meanwhile,145 serum samples were selected and compared with Roche electrochemiluminescence by linear regression analysis.Results A method for the detection of serum IL-6 based on acridine ester chemiluminescence immune quantitative analysis was established successfully.MES-BSA was selected as the dilution buffer of labeled antibody,0.2μg/ml was selected as the concentration of labeled antibody,and 50μl was selected as the sample addition amount.The LOB and LOD of the method were 0.2 pg/ml and 0.5 pg/ml,respectively,and the reportable range and the intermediate precision were 0.5~30000 pg/ml and less than 5.3%,respectively.When IL-6 concentration reached 200000 pg/ml,HOOK effect did not appear.In the concentration range of triglyceride(TG)30000μg/ml,hemoglobin(HGB)9000μg/ml,bilirubin(TBIL)330μg/ml and rheumatoid factor(RF)1500 IU/ml,the interfering substances did not affect on the detection results.Compared with the electrochemiluminescence method for IL-6 determination,the linear regression equation was Y=0.9802X-3.4879,r=0.9977,and the results of reagent determination by the two methods were highly correlated(P<0.05).Conclusion A method for the detection of serum IL-6

关 键 词：白细胞介素-6 吖啶酯 化学发光 免疫定量分析 

分 类 号：Q503[生物学—生物化学]