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

本研究的目的是建立(1)同时、(2)快速/简便和(3)鉴别的致泻菌检测方法。本研究取得了以下成果：(1)开发了一种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聚合物材料上化学固定抗体的新方法。(2)将上述方法成功地应用于两种副溶血性弧菌毒素和霍乱弧菌类似肠毒素(霍乱毒素)和产肠毒素大肠杆菌(不耐热肠毒素)的快速、简便、同时和区分检测。(3)进一步发展了一种通过在尼龙表面(聚合物材料)表面引入苯酚来选择性地将抗体铁片段化学固定在尼龙表面的新方法。这些将抗体化学固定在尼龙等聚合物材料上的新方法可广泛应用于各种传染病的免疫诊断试验。

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.