Mahony D E, Gilliatt E, Dawson S, Stockdale E, Lee S H
Department of Microbiology, Faculty of Medicine, Dalhousie University, Halifax, Nova Scotia, Canada.
Appl Environ Microbiol. 1989 Sep;55(9):2141-3. doi: 10.1128/aem.55.9.2141-2143.1989.
A rapid assay which measured the biological activity of Clostridium perfringens enterotoxin was developed. The method involved the rapid killing of Vero cells by enterotoxin produced by C. perfringens grown in Duncan and Strong sporulation medium. Serial dilutions of toxin were added to Vero cells either in suspension or grown as monolayers in wells of a 96-well cell tissue culture cluster plate. Vital staining of Vero cells with neutral red, followed by extraction of the dye, allowed toxin levels to be determined either visually or by optical density measurements with a micro-ELISA M580 computer program. The toxin produced was confirmed as different from the Vero toxin of Escherichia coli and the alpha and theta toxins of C. perfringens.
开发了一种快速检测产气荚膜梭菌肠毒素生物活性的方法。该方法利用在邓肯和斯特朗芽孢形成培养基中生长的产气荚膜梭菌产生的肠毒素快速杀死Vero细胞。将毒素的系列稀释液添加到悬浮培养的Vero细胞中，或添加到96孔细胞组织培养板孔中单层生长的Vero细胞中。用中性红对Vero细胞进行活细胞染色，然后提取染料，可通过目视或使用微型ELISA M580计算机程序进行光密度测量来确定毒素水平。所产生的毒素经证实不同于大肠杆菌的Vero毒素以及产气荚膜梭菌的α毒素和θ毒素。
