[食品中大肠杆菌O157多克隆抗体及双抗体酶联免疫吸附测定法的研究]

[Studies on the polyclonal antibody and the double-antibody enzyme-linked immunosorbent assay against E. coli O157 in food].

作者信息

Zhao Zhijing, Liu Xiumei

机构信息

National Institute of Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing 100050, China.

出版信息

Wei Sheng Yan Jiu. 2003 Nov;32(6):606-9.

PMID:14963916

Abstract

The polyclonal antibody (PcAb) for E. coli O157:H7 was available by intravenous injection with New Zealand rabbit with E. coli O157:H7 ATCC43895 strain. Together with the monoclonal antibody (McAb) 3A5 specific against E. coli O157 and O113:H21, a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed with the as the capture antibody and the McAb 3A5 as the detection antibody. The detection limits are 10(3)-10(4) cfu/ml in pure culture of E. coli O157 and without any cross reaction with the non-O157 strains. With 10 h enrichment procedure, E. coli O157:H7 recovered well from inoculated ground poultry meat and pasteurized milk at levels of 0.1 cfu/g(cfu/ml). The results demonstrated that the double-antibody sandwich ELISA is a sensitive and specific method for detecting E. coli O157 in food samples.

摘要

通过用大肠杆菌O157:H7 ATCC43895菌株对新西兰兔进行静脉注射获得了针对大肠杆菌O157:H7的多克隆抗体（PcAb）。与针对大肠杆菌O157和O113:H21的单克隆抗体（McAb）3A5一起，以该多克隆抗体为捕获抗体、McAb 3A5为检测抗体，建立了双抗体夹心酶联免疫吸附测定（ELISA）法。在大肠杆菌O157的纯培养物中，检测限为10³-10⁴ cfu/ml，且与非O157菌株无任何交叉反应。经过10小时的富集程序，在接种的禽肉碎和巴氏杀菌牛奶中，当大肠杆菌O157:H7的含量为0.1 cfu/g（cfu/ml）时能很好地被检出。结果表明，双抗体夹心ELISA是检测食品样品中大肠杆菌O157的灵敏且特异的方法。