单克隆抗体酶联免疫方法检测人血清胰岛素原及其应用  

Application of monoclonal antibody-based sandwich enzymeimmunoassay in detection of human serum proinsulin

作　　者：贾恩志[1] 李爱香[2] 徐振霞[2] 

机构地区：[1]南京医科大学第一附属医院心内科,江苏省南京市210029 [2]山东省济宁市第二人民医院急诊科,山东省济宁市272049

出　　处：《世界华人消化杂志》2005年第13期1562-1566,共5页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助课题;No.30400173~~

摘　　要：目的:建立灵敏、特异的检测人血清中胰岛素原的酶联免疫吸附试验并应用于临床与流行病学研究. 方法:选择2种mAb并引入生物素与亲和素放大系统来建立酶联免疫分析方法,一种抗C肽mAb结合到酶反应板上作为固相抗体,另一种生物素标记的抗胰岛素抗体作为液相抗体,将该检测方法应用于流行病学研究(n=1196). 结果:本法灵敏度0.83 pmol/L,与1 200 pmol/L胰岛素、3 960 pmol/L C肽无交叉反应,线性标准范围0.83-142 pmol/L.批内变异系数小于11.4%,批间变异系数小于11.2%.流行病学研究结果显示胰岛素原的含量与体重指数、腰围、收缩压、舒张压、胆固醇、三酰甘油、低密度脂蛋白胆固醇、危险因素数目呈正相关, 与高密度脂蛋白胆固醇呈负相关. 结论:本法适用于在临床研究及流行病学研究中检测人血清中胰岛素原的含量.β细胞功能受损与纤溶功能的紊乱可能是高胰岛素原与心血管危险因素关联的原因.AIM: To develop a highly sensitive and specific ELISA to detect the human serum proinsulin, and to explore the relationship between proinsulin and cardiovascular risk factors in a population-based epidemiological study. METHODS: The assay was based on two monoclonal antibodies, an anti-C-peptide antibody bound to a microtest plate and a biotin-labeled anti-insulin antibody. The biotin-avidin ELISA was used to measure the serum proinsulin of 1196 subjects who aged 35-59 in Pizhou, Jiangsu Province. RESULTS: With a detection limit of 0.83 pmol/L, the ELISA covered a standard linear range of 0.83-142 pmol/L, and the human insulin and human C-peptide did not cross-react at 1 200 and 3 960 pmol/L respectively. The inter-and inner-assay coefficient of variation of the ELISA were less than 11.4% and 11.2% respectively. The results of the population-based epidemiological study indicated that the concentration of proinsulin had significantly positive correlations with body mass index (BMI), waist, systolic blood pressure (SBP), diastolic blood pressure (DBP), total cholesterol (TCH), triglycerides (TG), low density lipopro-tein cholesterol (LDL-C), and cardiovascular risk factors clustering and negative association with high density lipo-protein cholesterol (HDL-C). CONCLUSION: The monoclonal-based BA-ELISA can be used to detect serum proinsulin in the clinical and epide-miological study. The mechanism underlying the relationship between proinsulin and cardiovascular risk factors may be related to the fibrinolysis and damages of β-cells.

关 键 词：胰岛素原 人血清 酶联免疫方法 单克隆抗体 检测 流行病学研究 低密度脂蛋白胆固醇 高密度脂蛋白胆固醇 mol/L 酶联免疫分析方法 酶联免疫吸附试验 心血管危险因素 抗胰岛素抗体 变异系数 生物素标记 放大系统 固相抗体 

分 类 号：R446.6[医药卫生—诊断学]