单克隆抗体检测分析S．typhiO，H，Vi抗原方法的建立及初步应用  

Establishment and Preliminary Application of Double-McAb Sandwich ELISA for Detection and Analysis of S.typhi O,H,and ViAntigens

作　　者：魏博[1] 彭静[1] 周学良[1] 

机构地区：[1]卫生部兰州生物制品研究所

出　　处：《单克隆抗体通讯》1995年第1期16-21,共6页

基　　金：甘肃省中青年科技基金

摘　　要：以Ｓ．ｔｙＰｈｉ全菌体免疫ＢＡＬＢ／ｃ小鼠，通过细胞融合技术，建立了分别针对Ｓ．ｔｙＰｈｉＯ９、Ｏ１２、Ｈｄ和Ｖｉ抗原的单克隆抗体（ＭＣＡｂ）杂交瘤共３０株。通过对ＭＣＡｂ的亚类、效价和表位测定等进行筛选，建立了可分别检测Ｓ．ｔｙＰｈｉ可溶性Ｏ、Ｈ和Ｖｉ抗原的双抗体夹心ＥＬＩＳＡ法。检测３种抗原成分的敏感度分别为：１５．６２５ｎｇ／ｍｌ，８ｎｇ／ｍｌ和０．３０５ｎｇ／ｍｌ。检测全菌体的敏感度为１０５～１０６个／ｍｌ。检测了疑似伤寒患者的血培养物１０４份，其中２７份三种抗原检测全部呈阳性，与细菌分离的结果完全相同，从而证实此２７例为Ｓ．ｔｙｐｈｉ感染。上述结果提示：此方法具有较高的敏感性及特异性。对Ｓ．ｔｙｐｈｉ抗原成分的检测分析，将可能应用于伤寒的早期诊断。Totally thirty murine hybridoma McAbs reactive with 09,012, Hd and Vi antigens ofS.typhi were obtained from Cell fusion by immunolising BALB/c mice with whole cellsof S.typhi.Several McAbs to different antigens were selected on the basis of detection ofimmunoglobulin subelass, antibody titer of and epitope. Double antibody sandwich ELISA was developed using IgG-class McAb as capture or probe antibody to detect O, H, and Vi antigens ofS. typhi.The sensitivities of detection for soluable S.typhi liPOPOlysaccharide(O antigen),flagellin (Hantigen), and Vi were 15.625 ng /ml,sng/ml and 0.305ng/ ma respectively.The ldriit of detectionfor whole bacteria was 105～106 cells/ml.By use of the assay O,H,and Vi antigens were detectedin 27 of the 104 hemoculture samples from patients.Positive samples were confirmed to be infectedwith S.typhi by bacteria isolation.The results indicate that the assay has high sensitivity andspecificity.The assays for detection and analysis of antigen comPOnents might be applied to the early,rapid diagnosis of typhoid fever.

关 键 词：伤寒杆菌 抗原 单克隆抗体 ELISA 

分 类 号：R378.23[医药卫生—病原生物学]