流式免疫微球技术定量检测人血清胰岛素方法学研究和评价  

Establishment and evaluation of a flow cytometric immune microsphere assay for quantitative detecting human serum insulin

作　　者：王占科[1,2] 何丹[3] 祝仲珍[2] 胡晓璐[4] 潘小清[2] 常津[1] 

机构地区：[1]天津大学,330072 [2]解放军第九四医院检验病理科,南昌330002 [3]江西省人民医院中心实验室,南昌330006 [4]南昌大学医学院,330006

出　　处：《免疫学杂志》2012年第4期341-343,348,共4页Immunological Journal

基　　金：国家科技支撑项目(2008BAI52B03)

摘　　要：目的建立流式免疫微球定量检测人血清胰岛素的方法并对其重复性实验和线性实验以及最低检测限进行评价。方法将兔抗人胰岛素单克隆抗体标记在微球表面制备免疫诊断微球,与血清中胰岛素结合后,加入荧光标记的鼠抗人胰岛素单克隆抗体,使微球表面呈现荧光,并通过流式细胞仪检测其平均荧光强度(mean fluorescent intensity,MFI)并记录。结果微球表面MFI与人血清胰岛素浓度呈正比,流式免疫微球技术检测同一标本的重复性实验变异系数以及最低检测限均低于酶联免疫吸附试验,流式免疫微球技术线性范围大于酶联免疫吸附实验。结论流式免疫诊断微球技术可用于人血清胰岛素定量检测,其测定重复性,线性范围和灵敏度均优于酶联免疫吸附试验。We aim to establish a novel flow cytometric immune microsphere assay（FCIMA） for quantitative detecting human serum insulin by flow cytometric immune microsphere assay（FCIMA） and study its repeatability,linearity,and lowest detectable limit（LDL）.Microsphere surface were labeled by rabbit against human insulin monoclonal antibody to prepare immune diagnosis microspheres,which could be combined with human insulin of serum,and appear fluorescent light by combining rat against human insulin monoclonal antibody labeled with FITC.The mean fluorescent intensity（MFI） of the surface of immune diagnosis microspheres were detected by flow cytometer（FC） and recorded.We found that the MFI of immune diagnosis microspheres were in direct proportion to the concentrations of human serum insulin.Moreover,both of the coefficient of variance（CV） of repeatability and the LDL of FCIMA were lower than those of ELISA,and the range of linearity of FCIMA was wider than that of ELISA.These results indicated that FCIMA can be used for quantitative determination of human serum insulin,which is better than ELISA in the repeatability,linearity range,and detection sensitivity.

关 键 词：流式免疫微球技术 平均荧光强度 定量测定 胰岛素 

分 类 号：R446.61[医药卫生—诊断学]