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Rapid Diagnosis of Campylobacter Infection by Latex Agglutination Using Monoclonal Antibodies.

使用单克隆抗体通过乳胶凝集快速诊断弯曲杆菌感染。

基本信息

批准号：
61571118
负责人：
NAKAMURA Yoshiko
金额：
$ 1.28万
依托单位：
Showa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1988
项目状态：
已结题

项目摘要

Campylobacter jejuni, the most frequent pathogens of acute gastroenteritis, usually requires two days for the isolation and identification. We made monoclonal antibodies(MoAb) against C. iejuni so as to search the frequencies of C. jejuni isolation and to establish the method for rapid diagnosis.1) The frequencies of pathogens isolated from feces were 36% in C. jejuni/ coli, 27% in Klebsiella oxytoca, 14% in enteropathogenic E. coli and 9% in Salmonella spp.2) Campylobacter coli shared 8% in all C. jejuni/coli.3) Eleven anti C. jejuni monoclonal antibodies were obtained by means of heat extracted C. jejuni C_1 strain. The epitopes reactive with these MoAb were polypeptides having 45kDa(6 strains) and 65kDa(5 strains) molecular masses on SDS-PAGE. The classes of MoAb were identified as IgM(2 strains) and IgG(9 strains).4) MoAb(H-194) sensitized latex agglutination(reversed passive latex agglutination, RPLA) was verified to be Campylobacter specific when examined by other pathogens of gastroenteritis.5) The sensitivity of the RPLA was 4x10^4 CFU/ml, as calculated from the plate-RPLA and colony formation unit of C. jejuni C strain.6) All clinically isolated 121 strains of C. jejuni and 11 strains of C. coli showed positive RPLA.7) There was a 96%(249 of 260) total agreement between RPLA and the standard culture method. Among 200 feces showed positive culture, 198 were identified as positive RPLA. Nine feces among 60 negative culture showed positive results.In summary, the RPLA appears to be useful for rapid detection(within 10 min) of Campylobacter. The method is sensitive, specific, simple to perform and requires no costly instrument.

空肠弯曲菌是急性胃肠炎最常见的病原菌，其分离鉴定通常需要2天。我们制备了抗C. iejuni，以搜索C. 1）空肠弯曲菌粪便分离率为36%;空肠/大肠杆菌、产酸克雷伯氏菌、肠致病性大肠埃希氏菌分别为27%、14%。大肠弯曲杆菌在所有C. 3）11株抗C.用热浸法提取空肠C.空肠C_1菌株。SDS-聚丙烯酰胺凝胶电泳结果表明，与这些单抗反应的抗原表位为分子量为45 kDa（6株）和65 kDa（5株）的多肽。4）用H-194致敏的乳胶凝集试验（RPLA）检测其他病原菌，结果表明RPLA对弯曲菌具有特异性。6）临床分离的121株C. jejuni和11株C. 7）RPLA与标准培养法的符合率为96%（249/260）。在200份培养阳性粪便中，198份被鉴定为RPLA阳性。在60份阴性粪便中有9份呈阳性结果。综上所述，RPLA可用于快速检测弯曲杆菌（10 min内）。该方法灵敏度高、特异性强、操作简单、不需要昂贵的仪器。