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

大肠埃希菌、肠炎沙门氏菌、副溶血性弧菌、空肠弯曲菌、金黄色葡萄球菌、蜡样芽孢杆菌、产气荚膜梭菌、肠出血性大肠埃希菌（verotoxin, VT+）属于食物中毒菌群。为了确定食物中毒的可能诊断，制备了针对这些细菌的单克隆抗体（mab）。单克隆抗体经酶联免疫吸附试验（ELISA）和其他免疫学分析证实。(1)抗vt单抗：建立了21个抗肠出血性大肠杆菌的克隆。5种单克隆抗体与VT1和VT2反应。(2)抗埃希氏菌线圈单抗：建立了13个克隆。(3)抗肠炎沙门氏菌单抗：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发生反应。(4)抗空肠弯曲杆菌单抗：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抗：建立6个克隆。四种单克隆抗体与一些肠毒素发生反应。(6)抗蜡样芽孢杆菌单抗：建立了2个克隆。一个单抗与蜡样芽孢杆菌和产气荚膜梭菌衍生的磷脂酶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开发免疫层析（IC）方法。此测试可能只需要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。使用单克隆抗体的IC快速、简单、特异、适用于多种样品，将使大多数实验室或研究所的食物中毒筛查和检测成为可能。

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.