Lyerly D M, Sullivan N M, Wilkins T D
J Clin Microbiol. 1983 Jan;17(1):72-8. doi: 10.1128/jcm.17.1.72-78.1983.
Antibodies against Clostridium difficile toxin A were purified by affinity chromatography from antiserum prepared against crude C. difficile toxin preparations. The affinity-purified antibody preparation was free of detectable amounts of antibodies to other C. difficile antigens, as demonstrated by crossed immunoelectrophoresis, and specifically neutralized the cytotoxicity of toxin A. An indirect enzyme-linked immunosorbent assay (ELISA) was subsequently developed using the antibody preparation for the specific detection of toxin A. The ELISA, which could detect 1 ng (5 ng/ml) of toxin A, was used to quantitate the toxin in the culture supernatant fluids of strains of C. difficile. The ELISA values for toxin A closely correlated with the toxin A and B cytotoxic titers of the supernatant fluids. In addition, toxin A was detected by ELISA in human fecal specimens from persons with antibiotic-associated colitis, demonstrating that this toxin is produced during C. difficile colitis.
用抗艰难梭菌毒素粗制品制备的抗血清，通过亲和层析法纯化抗艰难梭菌毒素A抗体。交叉免疫电泳表明，亲和纯化的抗体制剂不含可检测到的针对其他艰难梭菌抗原的抗体，且能特异性中和毒素A的细胞毒性。随后，利用该抗体制备了间接酶联免疫吸附测定法（ELISA），用于特异性检测毒素A。该ELISA法可检测1 ng（5 ng/ml）毒素A，用于定量艰难梭菌菌株培养上清液中的毒素。毒素A的ELISA值与上清液中毒素A和B的细胞毒性滴度密切相关。此外，在抗生素相关性结肠炎患者的人粪便标本中，通过ELISA检测到了毒素A，表明该毒素在艰难梭菌结肠炎期间产生。
