Hoy C S, Sesardic D
Division of Bacteriology, National Institute for Biological Standards and Control, South Mimms, Hertfordshire EN6 3QG, UK.
Toxicol In Vitro. 1994 Aug;8(4):693-5. doi: 10.1016/0887-2333(94)90045-0.
Subunit-specific antipeptide antibodies have been used to develop a dot blot immunoassay for detection of diphtheria toxin (DT) produced by toxigenic strains of Corynebacterium diphtheriae. A detection limit of 0.1-1.0 ng DT/ml was obtained. DT is cytotoxic to cultured Vero cells and cell viability if conveniently assessed using a tetrazolium dye, MTT. This has been used as the basis of a cytotoxicity assay for DT, which has a detection limit of 20 pg/ml of biologically active toxin. Sixty isolates of C. diphtheriae were assayed for toxigenicity. There was complete concordance between the Vero cell cytotoxicity assay and guinea pig intradermal assay in vivo, demonstrating the utility of the cytotoxicity assay.
亚单位特异性抗肽抗体已被用于开发一种斑点印迹免疫测定法，以检测产毒白喉棒状杆菌菌株产生的白喉毒素（DT）。获得的检测限为0.1 - 1.0 ng DT/ml。DT对培养的非洲绿猴肾细胞（Vero细胞）具有细胞毒性，并且如果方便地使用四唑盐染料MTT评估细胞活力。这已被用作DT细胞毒性测定的基础，该测定法对生物活性毒素的检测限为20 pg/ml。对白喉棒状杆菌的60个分离株进行了产毒性测定。Vero细胞细胞毒性测定与豚鼠体内皮内测定完全一致，证明了细胞毒性测定的实用性。
