Department of Microbiology, Institute of Biology, Jan Kochanowski University, Kielce, Poland.
Biochemistry (Mosc). 2011 Jul;76(7):851-61. doi: 10.1134/S0006297911070169.
The aim of this work was to serotype Proteus mirabilis urinary tract infection (UTI) strains based on chemically defined O-antigens with the use of two clinical collections from Sweden and Poland consisting of 99 and 24 UTI strains, respectively. A simple two-step serotyping scheme was proposed using enzyme immunoassay with heat-stable surface antigens of Proteus cells and immunoblotting with isolated lipopolysaccharides (LPSs). Using polyclonal anti-P. mirabilis rabbit antisera, 50 Swedish and 8 Polish strains were classified into serogroups O10, O38, O36, O30, O17, O23, O9, O40, O49, O27, O5, O13, O24, O14, and O33. From the Swedish strains, 10 belonged to serogroup O10 and five to each of serogroups O38, O36, and O9. Therefore, none of the O-serogroups was predominant. The majority of the serotyped clinical strains possess acidic O-antigens containing uronic acids and various acidic non-carbohydrate substituents. In immunoblotting, antisera cross-reacted with both O-antigen and core of LPSs. The core region of 19 LPSs bound a single serum, and that of 12 LPSs bound more than two sera. Following bioinformatic analysis of the available sequences, a molecular approach to the prediction of Proteus core oligosaccharide structures was proposed. The identification of the core type of P. mirabilis R110, derived from a serogroup O3 wild strain, using restriction fragments length polymorphism analysis of galacturonic acid transferase is shown as an example. In summary, the most frequent O-serogroups among P. mirabilis UTI stains were identified. The diversity of serological reactions of LPSs is useful for serotyping of P. mirabilis clinical isolates. A possible role of the acidic components of O-antigens in UTI is discussed.
本工作旨在基于化学定义的 O-抗原对奇异变形杆菌尿路感染(UTI)菌株进行血清型分型,为此使用了分别来自瑞典和波兰的两个临床菌株集,分别包含 99 株和 24 株 UTI 菌株。提出了一种简单的两步血清分型方案,使用酶免疫测定法结合变形杆菌细胞耐热表面抗原和免疫印迹法结合分离的脂多糖(LPS)。使用多克隆抗奇异变形杆菌兔抗血清,将 50 株瑞典菌株和 8 株波兰菌株分为 O10、O38、O36、O30、O17、O23、O9、O40、O49、O27、O5、O13、O24、O14 和 O33 血清群。在瑞典菌株中,有 10 株属于 O10 血清群,5 株属于 O38、O36 和 O9 血清群。因此,没有一个 O-血清群占主导地位。大多数经血清分型的临床菌株具有含有糖醛酸和各种酸性非碳水化合物取代基的酸性 O-抗原。在免疫印迹中,抗血清与 LPS 的 O-抗原和核心都发生交叉反应。19 个 LPS 的核心区域与单个血清结合,12 个 LPS 的核心区域与多个血清结合。在对现有序列进行生物信息学分析后,提出了一种预测奇异变形杆菌核心寡糖结构的分子方法。以源自 O3 野生株的奇异变形杆菌 R110 为例,使用半乳糖醛酸转移酶的限制片段长度多态性分析对其核心型进行鉴定。总之,确定了奇异变形杆菌 UTI 菌株中最常见的 O-血清群。LPS 血清学反应的多样性可用于奇异变形杆菌临床分离株的血清分型。还讨论了 O-抗原酸性成分在 UTI 中的可能作用。