一种新的无传染性的检测HBeAg抗HBe的单克隆酶联免疫吸附法(MonoLISA)的建立与应用  

USE OF A NON-INFECTIOUS MONOCLONAL ELISA(MonoLISA) FOR DETECTION OF HBeAg/ANTI-HBe

作　　者：詹美云[1] 苏丽娅[1] 张文英[1] 田瑞光[1] 汤少华[1] 张满仓 刘崇柏[1] 

机构地区：[1]中国预防医学科学院病毒学研究所

出　　处：《病毒学报》1991年第1期49-53,共5页Chinese Journal of Virology

基　　金：国家<七五>攻关课题

摘　　要：应用我室自1988年5月以来建立的抗乙型肝炎e抗原两个抗原决定簇的单克隆抗体,和在大肠杆菌中高效表达的乙型肝炎病毒e抗原,组装成检测HBeAg/抗HBe的酶联免疫试剂(简称Mo-noLISA)。测定了该试剂的特异性、敏感性和可重复性,并与用日本单克隆抗体和血清e抗原组装的类似的EIA试剂进行了比较。结果两种试剂检测92份乙肝病人血清e抗原和38份血清e抗体的总符合率,分别为97.8％和100％。用两种试剂同时滴定一份已知HBeAg阳往和一份抗-HBe阳性的血清,证明两者的敏感性无显著差异。重复性试验证实,该试剂的重复率为100％。由此证明,用抗-HBeMcAb和基因工程抗原组装的诊断试剂,特异性强,敏感性高,可重复性好,且无传染性。这不仅为诊断试剂原材料的更新提供了丰富的来源,而且将该类试剂提高到一个新水平。16 hybridomas that secret antibodies to HBeAg were established and used with HBeAg expressed in. E. coli to develop a specific, high sensitivity reagents for the detection of HBeAg/anti-HBe, as compared with Japanese McAb to HBeAg EIA kit ( supplied by Institute of Liver Disease, Beijing Medical College ) . McAb to HBeAg a determinant was used for coating wells, McAb to HBeAg β determinant was labelled with hoseradish peroxidase ( HRp ) , for testing HBeAg a sandwich method was used. For anti-HBe a competitive MonoLISA was used. The results showed that the HBeAg coincident rate was 97.8% ( Tab. 1) between Chinese MonoLISA and Japanese monoclonal EIA. The anti-HBe coincident rate was 100% (Tab.2). The sensitivity of MonoLISA for HBeAg was even higher than Japanese monoclonal EIA(Tab.3). For anti-HBe, there was no significant difference ( Tab. 4 ) . The repetition rates of MonoLISA for HBeAg and anti-HBe were 100%. Besides, this MonoLISA reagent for HBeAg/anti-HBe has seveal advantages. Firstly, it is non-infectious; secondly, both McAbs to HBeAg α or β determinants are used, either to coat wells or to label with HRP; thirdly, there is no effect wether HBeAg containing HBeAg or not is used the detection of anti-HBe.

关 键 词：单克隆抗体 检测 HBEAG MonoLISA 

分 类 号：R446.61[医药卫生—诊断学]