Department of Veterinary Sciences, Faculty of Veterinary Medicine, Ludwig-Maximilians-Universität München, Oberschleißheim, Germany.
Department of Biology I, Plant Development and Electron Microscopy, Biocenter Ludwig-Maximilians Universität München, Planegg-Martinsried, München, Germany.
PLoS One. 2022 Mar 17;17(3):e0265425. doi: 10.1371/journal.pone.0265425. eCollection 2022.
One of the multiple factors determining the onset of the diarrhoeal disease caused by enteropathogenic Bacillus cereus is the ability of the bacteria to actively move towards the site of infection. This ability depends on flagella, but it also varies widely between different strains. To gain more insights into these strain-specific variations, polyclonal rabbit antisera as well as monoclonal antibodies (mAbs) were generated in this study, which detected recombinant and natural B. cereus flagellin proteins in Western blots as well as in enzyme immunoassays (EIAs). Based on mAb 1A11 and HRP-labelled rabbit serum, a highly specific sandwich EIA was developed. Overall, it could be shown that strain-specific swimming motility correlates with the presence of flagella/flagellin titres obtained in EIAs. Interestingly, mAb 1A11, recognizing an epitope in the N-terminal region of the flagellin protein, proved to inhibit bacterial swimming motility, while the rabbit serum rather decreased growth of selected B. cereus strains. Altogether, powerful tools enabling the in-depth characterization of the strain-specific variations in B. cereus swimming motility were developed.
导致肠致病性蜡样芽孢杆菌引起的腹泻病的多个因素之一是细菌主动向感染部位移动的能力。这种能力取决于鞭毛,但不同菌株之间也有很大差异。为了更深入地了解这些菌株特异性的变异,本研究中产生了多克隆兔抗血清和单克隆抗体(mAb),这些抗体在 Western blot 以及酶免疫分析(EIA)中检测到重组和天然蜡样芽孢杆菌鞭毛蛋白。基于 mAb 1A11 和 HRP 标记的兔血清,开发了一种高度特异性的夹心 EIA。总的来说,可以表明菌株特异性的泳动能力与在 EIA 中获得的鞭毛/鞭毛蛋白滴度的存在相关。有趣的是,识别鞭毛蛋白 N 端区域表位的 mAb 1A11 被证明能抑制细菌的泳动能力,而兔血清则降低了所选蜡样芽孢杆菌菌株的生长。总之,开发了强大的工具,可以深入表征蜡样芽孢杆菌泳动能力的菌株特异性变异。