Development of rapid, simple and specific diagnosis using monoclonal antibodies to bacterial and viral food poisoning

使用单克隆抗体开发针对细菌和病毒食物中毒的快速、简单和特异性诊断

• 批准号: 16613009
• 负责人: KITAMOTO Noritoshi
• 金额: $ 2.43万
• 依托单位: University of Hyogo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16613009/
• 关键词: Monoclonal antibodies; Food poisoning; Specific diagnosis; ELISA method; Immunochromatography; Verotoxin; Enterotoxin; Heat-labile toxin; 病原大腸菌; 腸管出血性大腸菌; 黄色ブドウ球菌; サルモネラ; ノロウイルス

Escherichia coli, Salmonella enteritidis, Vibrio parahaemolyticus, Campylobacter jejuni, Staphylococcus aureus, Bacillus cereus, Clostridium perfringens, and enterohemorrhagic Escherichia coli (verotoxin, VT+) belong to the group of food poisoning bacteria. To establish the possible diagnosis for the food poisoning, monoclonal antibodies (MAbs) against these bacteria were produced. The MAbs was confirmed by an Enzyme-I inked immunosorbent assay (ELISA) test and other immunological analyses.(1) anti-VT MAb : 21 clones against enterohemorrhagic Escherichia coli were established. Five MAbs reacted with the VT1 and VT2.(2) anti-Escherichia coil MAb : 13 clones were established. One MAb may react with heat-1abiIe toxin (LT).(3) anti-Salmonella enteritidis MAb : 14 clones were established. However, these MAbs reacted with only strain used by immunogen.(4) anti-Campylobacter jejuni MAb : 5 clones were established. These MAbs reacted with some bacterial extract.(5) anti-Staphylococcus aureus MAb : 6 clones were established. Four MAbs reacted with some enterotoxins.(6) anti-Bacillus cereus MAb : 2 clones were established. One MAb reacted with phospholipase C derived from both Bacillus cereus and Clostridium perfringens.The MAbs against Vibrio parahaemolyticus and Clostridium perfringens were now under investigation.We will develop the inmunochromatography (IC) method using these MAbs. This test may require only 15 min to perform. It is important to prepare the diagnosis of food poisoning in an emergency. The IC using MAbs is rapid, simple, specific, applied for multiple samples, and will make it possible to screen for and detect food poisoning in most of laboratories or institutes.

大肠杆菌、沙门氏菌、副溶血性弧菌、空肠弯曲杆菌、金黄色葡萄球菌、蜡状芽孢杆菌、产气荚膜梭菌和肠出血性大肠杆菌（verotoxin，VT+）属于食物中毒菌群。为了建立食物中毒的可能诊断，产生了针对这些细菌的单克隆抗体（MAbs）。通过酶联免疫吸附测定（ELISA）试验和其他免疫学分析来确认单克隆抗体。(1)抗VT单克隆抗体：建立了21个抗肠出血性大肠杆菌的克隆。5株单抗与VT 1和VT 2反应。(2)抗大肠杆菌单克隆抗体：建立了13个克隆。一种单克隆抗体可与耐热毒素（LT）反应。(3)抗沙门氏菌单克隆抗体：建立了14个克隆。然而，这些单克隆抗体仅与免疫原使用的菌株反应。(4)抗空肠弯曲杆菌单克隆抗体：建立了5个克隆。这些单克隆抗体与一些细菌提取物反应。(5)抗金黄色葡萄球菌单克隆抗体：建立了6个克隆。四种单克隆抗体与一些肠毒素发生反应。(6)抗蜡样芽孢杆菌单克隆抗体：建立了2个克隆。其中一株单克隆抗体可与蜡样芽孢杆菌和产气荚膜梭菌的磷脂酶C反应，目前正在研究抗副溶血性弧菌和产气荚膜梭菌的单克隆抗体，我们将利用这些单克隆抗体建立免疫层析（IC）方法。该测试可能只需要15分钟即可完成。在紧急情况下准备食物中毒的诊断是很重要的。单克隆抗体免疫层析技术具有快速、简便、特异、适用于多样品等优点，将使大多数实验室或研究机构用于食物中毒的筛查和检测成为可能。

项目成果

Immunogenicity of a brominated protein and successive establishment of a monoclonal antibody to dihalogenated tyrosine

• DOI: 10.1016/j.freeradbiomed.2004.09.013
• 发表时间: 2005-01-01
• 期刊: FREE RADICAL BIOLOGY AND MEDICINE
• 影响因子: 7.4
• 作者: Kato, Y;Kawai, Y;Osawa, T
• 通讯作者: Osawa, T

Identification and quantification of N[epsilon]-(hexanoyl) lysine in human urine by liquid chromatography/tandem mass spectrometry.

通过液相色谱/串联质谱法鉴定和定量人尿中的 Nε-(己酰基)赖氨酸。

• 发表时间: 2004
• 期刊: Free Radical Biology and Medicine 37
• 作者: Kato Y;Yoshida A;Naito M;Kawai Y;Tsuji K;Kimura M;Kitamoto N;Osawa T.
• 通讯作者: Osawa T.

食品微生物検出法ノ最新技術

检测食品微生物的最新技术

• 发表时间: 2006
• 期刊: New Food Indusutry 48・3
• 作者: Kitta;K.;et al.;北元憲利
• 通讯作者: 北元憲利

Time-lag estimate between dietary intake and breast cancer mortality in Japan.

• 发表时间: 2007
• 期刊: Asia Pacific journal of clinical nutrition
• 影响因子: 1.3
• 作者: E. Harashima;Y. Nakagawa;G. Urata;K. Tsuji;M. Shirataka;Y. Matsumura

Quantification of advanced glycation end product and oxidative stress biomarkers in human urine.

人尿液中晚期糖基化终产物和氧化应激生物标志物的定量。

• 发表时间: 2005
• 期刊: Ann. N. Y. Acad. Sci. 1043
• 作者: Kato Y;Mizukawa N;Yano J;Hosokawa T;Yoshida A;Naito M;Kawai Y;Tsuji K;Kitamoto N;Osawa T.
• 通讯作者: Osawa T.