Development of rapid, differential and simultaneous detection methods for diarrheagenic bacteria

致腹泻细菌快速、差异、同步检测方法的开发

• 批准号: 06557019
• 负责人: HONDA Takeshi
• 金额: $ 3.9万
• 依托单位: Research Institute for Microbial Diseases, Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06557019/
• 关键词: Antibody; Chemical immobilization; Dirrheagenic bacteria; Rapid diagnosis; Vibrio parahaemolyticus; Vibrio cholerae; 毒素原性大腸菌; 下痢; ELISA; 迅速検出; DNAプローブ; 病原因子; 耐熱性溶血毒; ナイロン; 化学固定法

The purpose of this research is to develop the (1) simultaneous, (2) rapid/easy and (3) differential detection for diarrheagenic bacteria. Following results were obtained through the period of this research.(1) Development of a novel method to chemically immobilize antibody on polymeric materials such as nylon by using polyethylene imine and maleic anhydride methylvinyl ether copolymer.(2) Furthermore, the above method was successfully applied for the rapid, simple, simultaneous and differential detection of two Vibrio parahaemolyticus toxins and analogous enterotoxins of Vibrio cholerae (cholera toxin) and enterotoxigenic Escherichia coli (heat-labile enterotoxin).(3) We further developed a novel method for selective, chemical immobilization of Fe fragment of antibody onto nylon surface (polymeric materials) by introducing phenol on its surface, resulting in making Fab fragment of the antibody available.These novel methods of chemical immobilization of antibody onto polymeric materials such as nylon can be widely applied for immunological disgnostic tests of various infectious diseases.

本研究的目的是开发（1）同时，（2）快速/简单和（3）鉴别检测产肠杆菌。通过本研究取得了以下成果。(1)利用聚乙烯亚胺和马来酸酐甲基乙烯基醚共聚物在尼龙等高分子材料上化学固定抗体的新方法。(2)此外，上述方法已成功地应用于两种副溶血性弧菌毒素和类似的肠毒素的霍乱弧菌（霍乱毒素）和产肠毒素大肠杆菌（不耐热肠毒素）的快速，简单，同时和差异检测。(3)我们进一步发展了一种新的方法，通过在尼龙表面引入苯酚，将抗体的Fe片段选择性地化学固定在尼龙表面（聚合物材料）上，从而使抗体的Fab片段成为可利用的抗体，这些新的抗体化学固定在聚合物材料（如尼龙）上的方法可广泛应用于各种传染病的免疫诊断试验。

项目成果

Nagayama, K. et al.: "Use of alkaline phosphatase-conjugated oligounucleotide probe for the gene encoding the bungle-forming pilus of enteropathogenic." J. Clin. Microbiol.34(11). 2819-2821 (1996)

Nagayama, K. 等人：“使用碱性磷酸酶缀合的寡核苷酸探针来检测编码肠道致病菌菌毛的基因。”

鳥岡 要他: "医学のあゆみ" 細菌性腸管感染症と粘膜バリア, 122-129 (1996)

Kaname Torioka 等人：《医学史》细菌性肠道感染和粘膜屏障，122-129 (1996)

Koike,N.et al.: "Rapid and differential detection of two analogous enterotoxins of vibrio cholerae and enterotoxigenic Escherichia coli by a modified" FEMS Immun.Med.Microbiol.(in press).

Koike,N.等人：“通过改良的”FEMS Immun.Med.Microbiol.（正在印刷中）对霍乱弧菌和产肠毒素大肠杆菌的两种类似肠毒素进行快速和差异检测。

Xu,M.et al.: "Construction and characterization of an isogenic mutant of V.parahaemolyticus having a deletion inthe TRH gene." Journal of Bacteriology. 176. 4754-4760 (1994)

Xu,M.等人：“TRH 基因缺失的副溶血弧菌同基因突变体的构建和表征。”

Nagayama,K.et al.: "Correlation between Cell-associated mannose-sensitive hamagglutination by V.parahaemolyticus and adherence to a human colonic" FEMS Microbiology Letters. 120. 207-210 (1994)

Nagayama,K.等人：“副溶血弧菌引起的细胞相关甘露糖敏感血凝作用与人类结肠粘附之间的相关性”FEMS 微生物学快报。