Banno Y, Kobayashi T, Kono H, Watanabe K, Ueno K, Nozawa Y
Rev Infect Dis. 1984 Mar-Apr;6 Suppl 1:S11-20. doi: 10.1093/clinids/6.supplement_1.s11.
Two toxins were isolated from a toxigenic strain of Clostridium difficile. The toxins were purified by gel filtration and ion-exchange column chromatography to homogeneity as judged from polyacrylamide gel electrophoresis and were designated D-1 and D-2. Toxin D-1 was lethal for mice, increased vascular permeability, and induced fluid accumulation in ligated rabbit ileal loops, and toxin D-2 displayed cytotoxicity in HeLa cells with a minimum of 1 pg of toxin. The molecular weights of toxins D-1 and D-2, as estimated by gel filtration, ranged from 550,000 to 600,000 and from 450,000 to 500,000, respectively. These toxins were heat labile and inactivated by pronase and trypsin. Amino acid analyses of both toxins showed them to be of relatively similar composition. Antisera prepared against purified toxin D-1 neutralized all of the biologic activities of toxin D-1, whereas it did not affect any of the biologic activities of toxin D-2. A sensitive latex-agglutination immunoassay was developed for screening for C. difficile toxin D-1 in patients with pseudomembranous colitis.
从一株产毒素的艰难梭菌中分离出两种毒素。通过凝胶过滤和离子交换柱色谱法将毒素纯化至同质，这是根据聚丙烯酰胺凝胶电泳判断的，并将其命名为D-1和D-2。毒素D-1对小鼠具有致死性，可增加血管通透性，并在结扎的兔回肠袢中诱导液体蓄积，而毒素D-2在HeLa细胞中显示出细胞毒性，最低只需1 pg毒素。通过凝胶过滤估计，毒素D-1和D-2的分子量分别为550,000至600,000和450,000至500,000。这些毒素对热不稳定，可被链霉蛋白酶和胰蛋白酶灭活。两种毒素的氨基酸分析表明它们的组成相对相似。针对纯化的毒素D-1制备的抗血清可中和毒素D-1的所有生物学活性，而对毒素D-2的任何生物学活性均无影响。开发了一种灵敏的乳胶凝集免疫测定法，用于筛查假膜性结肠炎患者中的艰难梭菌毒素D-1。
