层粘连蛋白时间分辨免疫分析方法的建立  

The Establishment of Time-resolved Fluoroimmunoassay for Human Laminin

作　　者：黄飚[1] 裴豪[2] 朱岚[1] 陈铁河[3] 蒋祥虎[2] 葛东波 

机构地区：[1]江苏省原子医学研究所,江苏无锡214063 [2]无锡市传染病医院检验科,江苏无锡214005 [3]上海海研医学生物技术中心,上海200433

出　　处：《标记免疫分析与临床》2007年第2期97-99,共3页Labeled Immunoassays and Clinical Medicine

基　　金：江苏省社会发展基金资助项目(BS2004009)

摘　　要：采用时间分辨荧光免疫分析（TRFIA）技术建立高灵敏的层粘连蛋白（LN）的快速的全自动检测方法.以羊抗鼠IgG抗体包被板,采用抗人LN单克隆抗体（McAb）、Eu3＋标记人LN和以β-二酮体为主的增强液建立竞争LN-TRFIA,数据采用双对数函数处理程序处理.结果表明LN的标记率为11.5 Eu^3＋/LN,方法的批内和批间CV分别为3.9%和6.7%,平均回收率为91.2%,灵敏度为5μg/L,可测范围为5～1000μg/L,ED20、ED50和ED80分别为723.1μg/L、183.4μg/L和56.95μg/L.临床结果与RIA结果相符.本文建立的LN-TRFIA灵敏、稳定,可全自动操作,有很好的应用前景.TRFIA for human laminin （LN） was developed using polyclonal antibody of rabbit anti-mouse coated on wells, antibody of mouse anti-human laminin as first antibody, laminin labeled with Eu^3＋ and fluorescent enhancement solution which mainly contained 2-naphthoyltrifluoroacetone. The results showed that the labeling yield of laminin with Eu^3＋ was 11.5 Eu^3＋/ LN. The detection range of LN was from 5 to 1000μg/L, the correlation coefficient of standard curve was more than 0. 935, the lowest detection level was 5μg/L, the average recovery rate was 91.2%, the intraassay CV was 3.9% and the interassay CV was 6.7%. The ED25, ED50 and ED80 was 723. 1μg/L, 183.4 μg/L and 56.95μg/L separately. The detected results with tis TRFIA closely corresponded to those of RIA. The TRFIA for human laminin developed in this study is reliable, sensitive, stable and could meet the need of clinical application.

关 键 词：时间分辨荧光免疫分析 层粘连蛋白 全自动检测方法 

分 类 号：R446.6[医药卫生—诊断学]