一步夹心法检测HBsAg的ELISA技术

ONE STEP INCUBATION ELISA FOR THE DETECTION OF HBsAg

本文介绍一种应用单克隆抗-HBs作为试剂,检测HBsAg的高灵敏度ELISA技术.单克隆抗-HBs的H3F5与D2H5株分别用作固相包被抗体与酶标记抗体.待检血清标本与酶标记抗体混合同期培育,45°C1h.洗板后加入底物(TMB)显色.灵敏度水平达到1～3ngHBsAg/ml(T/N值2.1N5.O).对32份已知HBsAg阴性(RIA)的正常人血清,本法无1例阳性.已知HBsAg阳性(RIA)携带者血清269份,用本法检测全部阳性.1000例各类人群血清,本法与RIA法平行测试,结果一致率达97.6%.

A new technique for the detection of HBsAg by one step incubation ELISA was developed.Two strains of monoclonal hybridoma cell(D2H5 and H3F5)were selected for the production of monoclonal anti-HBs.and the antibodies were used respectively for conjugation with peroxidase or for Coatring the solid phase in this;ELISA system.Procedure of the technique is described.The sensitivity level of this ELISA system was up to 1 to 3 ng HBsAg/ml with P/N ratio 2.1 to 5.0.32 HBsAg negative samples wero rested by this ELISA system repealedly,no false positive result was found.269 HBsAg carrier sera were detecfed parallel]y by both RIA and this ELISA system by different technicians, and the results were all found positive.1000 samples from the population in Guangzhou area were defermined by RIA and this ELISA system by two groups of technicians,the results were highly comparative.The authors hold tnat this assay is considered as a technique of high sensitity,simplicity and specificity.