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用于直接检测和定量产肠毒素大肠杆菌中F41和K99菌毛抗原的单克隆抗体被动血凝试验和捕获酶联免疫吸附测定

Monoclonal antibody passive hemagglutination and capture enzyme-linked immunosorbent assays for direct detection and quantitation of F41 and K99 fimbrial antigens in enterotoxigenic Escherichia coli.

作者信息

Raybould T J, Crouch C F, Acres S D

出版信息

J Clin Microbiol. 1987 Feb;25(2):278-84. doi: 10.1128/jcm.25.2.278-284.1987.

Abstract

Production of diarrhea in neonatal calves by enterotoxigenic Escherichia coli depends on its ability to attach to the epithelial cells of the intestine via surface adhesins called pili or fimbriae and to secrete enterotoxins. The most important of these fimbriae are designated K99 and F41. We produced and characterized a murine monoclonal antibody specific to F41. This monoclonal antibody and a K99-specific monoclonal antibody were used to develop sensitive and specific passive hemagglutination and capture enzyme-linked immunosorbent assays (ELISAs) for detection and quantitation of F41 and K99 antigens in E. coli cultures and culture supernatants. The capture ELISA systems exhibited excellent sensitivity and specificity, whereas the passive hemagglutination systems appeared to be oversensitive. The ability of the capture ELISAs to detect K99 and F41 fimbrial antigens in fecal specimens from calves was evaluated. Fimbrial antigens were detected in six of six specimens from scouring calves but not in four of four specimens from nonscouring calves.

摘要

产肠毒素大肠杆菌导致新生犊牛腹泻,这取决于该菌通过称为菌毛的表面黏附素附着于肠上皮细胞以及分泌肠毒素的能力。其中最重要的菌毛被命名为K99和F41。我们制备并鉴定了一种对F41特异的鼠单克隆抗体。该单克隆抗体和一种对K99特异的单克隆抗体被用于开发灵敏且特异的被动血凝试验以及捕获酶联免疫吸附测定(ELISA),以检测和定量大肠杆菌培养物及培养上清液中的F41和K99抗原。捕获ELISA系统展现出极佳的灵敏度和特异性,而被动血凝系统似乎过于敏感。评估了捕获ELISA检测犊牛粪便标本中K99和F41菌毛抗原的能力。在腹泻犊牛的6份标本中均检测到菌毛抗原,而在未腹泻犊牛的4份标本中均未检测到。

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