Ecological Studies on Food-Bacteria from Food and Environments

食品和环境中食品细菌的生态学研究

基本信息

批准号：
09460146
负责人：
UEMURA Takashi
金额：
$ 5.57万
依托单位：
Coolage of Agriculture, Osaka Prefecture University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

(1) To develop a rapid and specific method to detect and/or identify enterohemorrhagic Escberichia coli O 157 : H7, two mouse monoclonal antibodies (MCA) were prepared. Specificities of these two MAbs (1D9 and 3E8) were determined by flow cytometry method (FCM). 3E8 and 1D9 were found to react with E.coli O157 : H7, Citrobacter freundii and Salmonella group N (O : 30), but not with E bermannii With a mixture containing strains of E coil O157 : H7 and E coli O6 : Hl, two different peaks appeared in FCM with MCA, whereas a single peak appeared with polyclonal rabbit antiserum. From these findings, FCM with MCA is suggested to be a rapid, specific, and useful method to detect and identify strain(s) of E celi O157 : H7 in food ingredients (Ref.#1).(2) FCM with fluorescent-labeled anti-CPE antibody was applied to develop a rapid, specific, and convenient method to detect enterotoxin (CPE) exposed on the surface of spores of Clostridium perfringens. The results obtained indicate that FCM can specifically detect CPE exposed on C perfringens spores for a short time. Thus, FCM is found to be a rapid, specific, and convenient assay method for detection of CPE exposed on C perfringens spores, suggesting that it will be hopefully useful to diagnose food poisoning caused by C perfringens (Ref.#2).(3) Sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative estimation of C perfringens enterotoxin (CPE) using* monoclonal* and* polyclonal* antibodies* as* capturing* and* detecting* antibodies* respectively.* The dose- dependent relationship between absorbance at 405 nm and concentration of purified CPE was obtained over the range of 0.5 ng/ml-8.0 ? g/ml. The sandwich ELISA is found to detect crude CPE in culture and CPE in 10% fecal extracts. This method is a convenient, rapid and sensitive for specific detection of GPE(Ref#3).

（1）制定了一种快速而特定的方法来检测和/或识别肠hap虫大肠杆菌O 157：H7，准备了两种小鼠单克隆抗体（MCA）。通过流式细胞仪法（FCM）确定这两个mAb（1d9和3e8）的特异性。 3E8和1d9被发现与e.coli O157：H7，柠檬酸杆菌和沙门氏菌N（O：30）反应，但与e bermannii没有与含有E Coil O157：H7和E Coli O6和E Coli O6：HL的混合物的E Bermannii反应，在McA中出现了两种不同的峰。 From these findings, FCM with MCA is suggested to be a rapid, specific, and useful method to detect and identify strain(s) of E celi O157 : H7 in food ingredients (Ref.#1).(2) FCM with fluorescent-labeled anti-CPE antibody was applied to develop a rapid, specific, and convenient method to detect enterotoxin (CPE) exposed on the surface of spores of Clostridium灌注。获得的结果表明，FCM可以在短时间内特异性检测在C刺激孢子上暴露的CPE。因此，发现FCM是一种快速，具体且方便的测定方法，用于检测在C灌注孢子上暴露于CPE的CPE，这表明它将有希望可以诊断由C灌注环境引起的食物中毒是有用的（参考文献2）。（3）使用C酶 - 酶 - 酶 - 链接的免疫吸收分析（ELISA）的量子量（ELISA）是使用量子的量化估计的（ELISA）是有用的。单克隆*和*多克隆*抗体* AS*捕获*和*分别检测*抗体*。*在405 nm处的吸光度与纯化CPE浓度之间的剂量相关关系是在0.5 ng/ml-8.0的0.5 ng/ml-8.0中获得的？ g/ml。发现三明治ELISA可检测培养物中的粗糙CPE，在10％的粪便提取物中检测到CPE。对于特异性检测GPE是一种方便，快速且敏感的（参考＃3）。