流动注射协同增强化学发光免疫分析法测定胰岛素  

Flow injection immunoassay for insulin using a new synergistic enhanced chemiluminescence

作　　者：骆建新[1] 张晓[1] 杨秀岑[2] 伍莉萍[2] 邱岳[1] 宋海星[1] 

机构地区：[1]第三军医大学成都军医学院医学检验教研室,成都610083 [2]华西医科大学药学院无机教研室

出　　处：《中华检验医学杂志》2001年第4期233-235,共3页Chinese Journal of Laboratory Medicine

基　　金：卫生部科学研究基金课题 (9412 19) ;四川省卫生厅科学研究基金课题 (0 0 0 2 2 0 )

摘　　要：目的　建立一种测定胰岛素的非放射性免疫分析新方法。方法　将流动注射分析法、免疫分析法和化学发光分析法集为一体 ,以辣根过氧化物酶标记抗体 ,四苯硼钠与对苯苯酚协同增强的鲁米诺 过氧化氢发光混合液作为发光体系。根据固相夹心免疫分析原理 ,建立流动注射协同增强化学发光免疫分析法测定胰岛素。结果　在 0 .15～ 6 0 0 0 μU范围内 ,胰岛素量与发光强度有良好的线性关系 ,r=0 .9972 (P <0 .0 1) ,检测限为 0 .0 42 μU ;精密度为3.36 %～ 11.88% ,回收率为 92 .13%～97.87% ;免疫柱可在 1个月内反复使用 2 0 0次以上 ;本法测定胰腺提取液试样的结果 ,与放射免疫分析法的测定结果基本一致。结论　建立的胰岛素测定方法快速灵敏、特异性强、重现性好、操作简便、易于自动化 ,并且无放射性污染 。Objective To establish a new non radioimmunoassay for detecting insulin. Methods We put together the techniques of chemiluminescent analysis, flow injection analysis and immunoassay and established a method,flow injection chemiluminescent immunoassay, for insulin determination using sandwhich procedure with the horseradish peroxdase (HRP) as label of antibody and using the detection system of synergistic enhanced chemiluminescence of para phenylpenol and sodium tetraphenylborate with HRP catalysed oxidation of luminol. Results Satisfactory results were obtained in the dynamic range 0.15～60.00 μU with a relative coefficient of 0 997 2 ( P <0.01), a detection limit of 0.042 μU and precisions of 3.36%～11.88%. The recovery of the assay was 92.13%～97.87% for swine insulin. The lifetime of the immuo column was long enough, one cloumn was applied for more than 200 determinations in one month without significant loss of activity.The established method was used to determine swine insulin in the solutions extracted from frozen swine pancreas, and the results accorded with those of radioimmunoaasay. Conclusion Owing to its high speed, high sensitivity, high selectivity, and high precision, simplification of operation, ease for automization, without radioactive waste especially,the method established for determination of insulin, various potential applications.

关 键 词：流动注射分析 免疫测定 化学发光 胰岛素 固相夹心免疫分析 

分 类 号：R446.6[医药卫生—诊断学]