Kurazono H, Yamasaki S, Takeda Y
Department of Microbiology, Faculty of Medicine, Kyoto University.
Nihon Rinsho. 1995 Sep;53(9):2296-300.
A highly sensitive bead-enzyme-linked immunosorbent assay to detect bacterial protein toxins was developed. Fab' of anti-toxin IgG was conjugated with horseradish peroxidase by the maleimide method and tetramethylbenzidine was used as a substrate. As the solid phase, a 6.5 mm diameter polystyrene bead was used and this was coated with the anti-toxin IgG. The sensitivities of the bead-ELISA for various bacterial protein toxins were as follows: less than 40 pg/ml for cholera enterotoxin (CT), less than 20 pg/ml for VT1 and less than 6 pg/ml for VT2 of enterohemorrhagic Escherichia coli. The bead ELISA was evaluated for direct detection of CT from stool specimens of patients with acute secretory diarrhea. Of the 75 stool samples examined, 59 yielded biochemically and serologically confirmed strains of Vibrio cholerae O1. The bead ELISA was positive for CT in stool supernatants in 50 (84.7%) of the 59 samples from which V. cholerae O1 was isolated. In addition, the bead ELISA was positive for three stool specimens which were negative by culture. These data indicate that the bead ELISA is a sensitive and simple method for direct detection of CT in nonsterile stool samples.
开发了一种用于检测细菌蛋白毒素的高灵敏度珠式酶联免疫吸附测定法。通过马来酰亚胺法将抗毒素IgG的Fab'与辣根过氧化物酶偶联，并使用四甲基联苯胺作为底物。作为固相，使用直径6.5毫米的聚苯乙烯珠，并将其用抗毒素IgG包被。该珠式酶联免疫吸附测定法对各种细菌蛋白毒素的灵敏度如下：霍乱肠毒素（CT）小于40 pg/ml，肠出血性大肠杆菌的VT1小于20 pg/ml，VT2小于6 pg/ml。对该珠式酶联免疫吸附测定法进行了评估，以直接检测急性分泌性腹泻患者粪便标本中的CT。在所检查的75份粪便样本中，59份产生了经生化和血清学确认的霍乱弧菌O1菌株。在分离出霍乱弧菌O1的59份样本中的50份（84.7%）粪便上清液中，该珠式酶联免疫吸附测定法对CT呈阳性。此外，对于3份培养结果为阴性的粪便标本，该珠式酶联免疫吸附测定法也呈阳性。这些数据表明，该珠式酶联免疫吸附测定法是一种用于直接检测非无菌粪便样本中CT的灵敏且简单的方法。
