Welch D F, Menge S K, Matsen J M
J Clin Microbiol. 1980 May;11(5):470-3. doi: 10.1128/jcm.11.5.470-473.1980.
A counterimmunoelectrophoresis (CIE) technique which reacted positively with culture filtrates of Clostridium difficile was developed and compared with a cytotoxicity assay in human embryonic lung cell cultures. CIE, employing C. sordellii antitoxin, detected 17 of 17 C. difficile strains. Of those positive by CIE, 13 were cytotoxic in cell culture. Fourteen Clostridium species other than C. difficile, C. sordellii, and C. bifermentans were negative by CIE. C. sordellii and C. bifermentans gave positive CIE results but were not cytotoxic. Similar sensitivity of toxin detection was observed for both methods. Optimal conditions for performing CIE included use of 48-h chopped meat-glucose broth cultures as the antigen source, use of a 10x-concentrated U.S. Standard C. sordillii antitoxin, and electrophoresis for 1.5 h in 0.05 M tris(hydroxymethyl)aminomethane-barbital-sodium barbital, pH 8.8, at a constant current of 6 mA/slide. CIE appears to be a suitable alternative to the cytotoxicity assay and may serve as a means for presumptive identification of C. difficile.
一种与艰难梭菌培养滤液呈阳性反应的对流免疫电泳（CIE）技术被开发出来，并与人类胚胎肺细胞培养中的细胞毒性测定法进行了比较。采用索氏梭菌抗毒素的CIE检测出17株艰难梭菌中的17株。在CIE呈阳性的菌株中，有13株在细胞培养中具有细胞毒性。除艰难梭菌、索氏梭菌和双发酵梭菌外的14种梭菌属菌株经CIE检测为阴性。索氏梭菌和双发酵梭菌CIE结果呈阳性，但无细胞毒性。两种方法在毒素检测方面观察到相似的灵敏度。进行CIE的最佳条件包括使用48小时的碎肉-葡萄糖肉汤培养物作为抗原来源，使用10倍浓缩的美国标准索氏梭菌抗毒素，以及在0.05M三（羟甲基）氨基甲烷-巴比妥钠-巴比妥缓冲液（pH 8.8）中以6mA/载玻片的恒定电流电泳1.5小时。CIE似乎是细胞毒性测定法的一种合适替代方法，可作为艰难梭菌初步鉴定的手段。
