Development of monoclonal antibody against pathogenic fungus of red rot disease of Porphyra and its application for the early detection of the disease.

1.单克隆抗体的批量生产。用紫菜红腐病病原菌皮霉对小鼠进行免疫。在RPMI 1640培养基中培养杂交瘤，获得大量单克隆抗体。抗体滴度高，足以检测出毕氏菌。该单克隆抗体对来自爱知县和佐贺县的毕氏菌有反应，而对来自宫城县的毕氏菌无反应。该菌与陆生植物和藻类中的另一种皮霉均无交叉反应。海水中游动孢子出现的连续观察。：在九月至二月期间，我们每星期在紫菜养殖场抽取三公升海水一次或两次。在1月初至2月中旬观察到游动孢子。《zoospore》的发行高峰出现在2月初。最高游动孢子数为1184个/l。紫菜养殖网的早期对策。： 1月初在渔网上养殖的紫菜，虽然在海水中已检出游动孢子，但仍未感染红腐病。这种紫菜被收获并用于生产干燥的“紫菜”作为测试样本。作为对照，用染疫后的紫菜。生产的干紫菜的质量。用早期平生紫菜生产的“紫菜”干比对照高一个等级。早期计划的“紫菜”的投标价格比对照组高出约16%。对照紫菜提取物中，不仅谷氨酸、甘氨酸、丙氨酸等风味氨基酸含量，而且总游离氨基酸含量均比对照高50 ~ 60%。因此，利用单克隆抗体早期检测红腐病，降低红腐病发病率成为可能。

1.Mass production of the monoclonal antibody. : Pythium sp., the causative organism of laver red rot, was used to immunize mice. Hybridoma was cultured in RPMI 1640 medium and large quantity of the monoclonal antibody was obtained. The titer of the antibody was high enough to detect the Pythium. The monoclonal antibody reacted to the Pythium sp. from Aichi and Saga prefectures but not to that from Miyagi prefecture. No cross-reactivity was found against another Pythium from land plants and alage.2.Continuous observation of the appearance of zoospore in seawater. : Three liters of seawater were collected from Porphyra cultivation farms once or twice a week from September to February. Zoospore were observed from early January to middle February. The peak of the release of zoospore was in the beginning of February. The highest number of zoospores was 1184/litter.3.An early counterplan for Porphyra cultivation nets. : The Porphyra cultured on cultivation nets in early January was found still not infected with red rot disease, though zoospore were detected already in seawater. This Porphyra was harvested and used to produce dried 'Nori' as a test sample. As a control, Porphyra infected with the disease then after was used.4.Quality of dried 'Nori' produced. : The grade of dried 'Nori' produced from early counterplaned Porphyra was one grade higher than that of the control. The price tendered for the early counterplaned 'Nori' was about 16% higher than that of the control. In the extractives of counterplaned 'Nori', not only the levels of taste amino acids such as glutamic acid, glycine and alanine but also that of total free amino acid were about 50-60% higher than that of control.Therefore, early detection of red rot disease by means of monoclonal antibody and reducing the incidence of red rot disease became possible.