Department of Microbiological Surveillance and Research, Statens Serum Institut, 2300 Copenhagen S, Denmark.
J Med Microbiol. 2012 Mar;61(Pt 3):317-322. doi: 10.1099/jmm.0.036350-0. Epub 2011 Oct 20.
The ability of bacterial pathogens to express different fimbrial adhesins plays a significant role in virulence. Thus, specific detection of fimbrial expression is an important task in virulence characterization and epidemiological studies. Most clinical Klebsiella pneumoniae isolates express type 1 and type 3 fimbriae, which are characterized by mediation of mannose-sensitive agglutination of yeast cells and agglutination of tannic acid-treated ox red blood cells (RBCs), respectively. It has been observed that K. pneumoniae isolates agglutinate yeast cells and commercially available sheep RBCs in a mannose-resistant manner. Thus, this study was initiated to identify the adhesin involved. Screening of a mutant library surprisingly revealed that the mannose-resistant agglutination of yeast and sheep RBCs was mediated by type 3 fimbriae. Specific detection of type 1 fimbriae expression in K. pneumoniae was feasible only by the use of guinea pig RBCs. This was further verified by the use of isogenic fimbriae mutants and by cloning and expressing K. pneumoniae fimbrial gene clusters in Escherichia coli. Yeast agglutination assays are commonly used to detect type 1 fimbriae expression but should not be used for bacterial species able to express type 3 fimbriae. For these species, the use of guinea pig blood for specific type 1 fimbriae detection is essential. The use of commercially available sheep RBCs or yeast is an easy alternative to traditional methods to detect type 3 fimbriae expression. Easy and specific detection of expression of type 1 and type 3 fimbriae is essential in the continuous characterization of these important adhesive virulence factors present in members of the Enterobacteriaceae.
细菌病原体表达不同菌毛黏附素的能力在毒力中起着重要作用。因此,菌毛表达的特异性检测是毒力特征和流行病学研究的重要任务。大多数临床分离的肺炎克雷伯菌表达 1 型和 3 型菌毛,它们分别通过介导酵母细胞甘露糖敏感凝集和鞣酸处理的牛红细胞(RBC)凝集来表征。已经观察到肺炎克雷伯菌分离株以甘露糖抗性方式凝集酵母细胞和市售绵羊 RBC。因此,本研究旨在鉴定涉及的黏附素。突变文库的筛选出人意料地表明,酵母和绵羊 RBC 的甘露糖抗性凝集是由 3 型菌毛介导的。只有使用豚鼠 RBC 才能实现对肺炎克雷伯菌 1 型菌毛表达的特异性检测。这通过使用同源性菌毛突变体和在大肠杆菌中克隆和表达肺炎克雷伯菌菌毛基因簇进一步得到证实。酵母凝集试验常用于检测 1 型菌毛表达,但不应用于能够表达 3 型菌毛的细菌。对于这些物种,使用豚鼠血液进行特异性 1 型菌毛检测是必不可少的。使用市售绵羊 RBC 或酵母是替代传统方法检测 3 型菌毛表达的简单方法。易于检测 1 型和 3 型菌毛的表达对于不断表征肠杆菌科成员中这些重要的黏附性毒力因子至关重要。