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Updated: May 28, 2026

Purification and Visualization of Lipopolysaccharide from Gram-negative Bacteria by Hot Aqueous-phenol Extraction
Published on: May 28, 2012
Serotyping of Proteus mirabilis clinical strains based on lipopolysaccharide O-polysaccharide and core
W Kaca1, J Glenska, L Lechowicz
1Department of Microbiology, Institute of Biology, Jan Kochanowski University, Kielce, Poland. wieslaw.kaca@ujk.edu.pl
Abstract:
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.
Insights
This study serotyped Proteus mirabilis urinary tract infection strains using enzyme immunoassay and immunoblotting. Diverse O-serogroups were identified, highlighting the utility of lipopolysaccharide diversity for clinical isolate serotyping.
Area of Science:
- Microbiology and Immunology
- Molecular Biology
- Bioinformatics
Background:
- Proteus mirabilis is a significant cause of urinary tract infections (UTIs).
- Accurate serotyping of P. mirabilis strains is crucial for understanding infection epidemiology and pathogenesis.
- Existing serotyping methods require refinement for clinical applications.
Purpose of the Study:
- To serotype clinical Proteus mirabilis urinary tract infection strains based on O-antigens.
- To develop a simple, two-step serotyping scheme for P. mirabilis.
- To explore the structural diversity of lipopolysaccharides (LPSs) and their potential role in UTIs.
Main Methods:
- Utilized enzyme immunoassay with heat-stable surface antigens and immunoblotting with isolated lipopolysaccharides (LPSs).
- Employed polyclonal anti-P. mirabilis rabbit antisera for classification.
- Conducted bioinformatic analysis for predicting core oligosaccharide structures and restriction fragments length polymorphism analysis.
Main Results:
- Successfully classified 50 Swedish and 8 Polish P. mirabilis UTI strains into multiple O-serogroups (e.g., O10, O38, O36, O9).
- Observed no predominant O-serogroup, indicating significant strain diversity.
- Revealed that most strains possess acidic O-antigens and identified cross-reactivity in LPS core regions.
Conclusions:
- Established a reliable serotyping scheme for P. mirabilis clinical isolates based on LPS diversity.
- The identified O-serogroups provide valuable epidemiological data for P. mirabilis UTIs.
- Acidic components of O-antigens may play a role in UTI pathogenesis.
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