Identification and differentiation of Taylorella equigenitalis and Taylorella asinigenitalis by lipopolysaccharide O-antigen serology using monoclonal antibodies.
Abstract: Lipopolysaccharides (LPSs) from Taylorella equigenitalis, the causative agent of contagious equine metritis, and T. asinigenitalis were compared by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Lipopolysaccharide profiles of 11 T. equigenitalis strains were similar, but different from the profiles of 3 T. asinigenitalis strains, and the profiles of 2 T. asinigenitalis strains were similar to each other. The serological specificities of the LPSs from these 14 strains were examined by immunoblotting and enzyme-linked immunosorbent assay with monoclonal antibodies (MAbs) to the LPSs of the T. equigenitalis and T. asinigenitalis type strains and T. asinigenitalis strain 2329-98. A MAb to T. equigenitalis LPS O-polysaccharide (O-PS) (M2560) reacted with LPSs from all T. equigenitalis strains but did not react with LPSs from the 3 T. asinigenitalis strains or with 43 non-Taylorella bacteria. Three MAbs to the T. asinigenitalis type strain LPS O-PS or core epitopes (M2974, M2982, M3000) reacted with the homologous strain and T. asinigenitalis strain Bd 3751/05, but not with any of the other bacteria. Five MAbs to T. asinigenitalis 2329-98 LPS O-PS or core epitopes (M2904, M2907, M2910, M2923, M2929) reacted only with this strain. Proton nuclear magnetic resonance spectra of the O-PSs of the type strains of T. equigenitalis and T. asinigenitalis provided fingerprint identification and differentiation of these 2 organisms. The serological results were consistent with our previous finding that the O-antigen of the type strain of T. equigenitalis, being a linear polymer of disaccharide repeating [-->4)-alpha-L-GulpNAc3NAcA-(1-->4)-beta-D-ManpNAc3NAcA-(1-->] units, differs from that of the T. asinigenitalis O-antigen polymer that is composed of repeating [-->3)-beta-D-QuipNAc4NAc-(1-->3)-beta-D-GlcpNAmA-(1-->] units. Lipopolysaccharide O-PS could be a specific marker for identification and differentiation of T. equigenitalis and T. asinigenitalis, and provide the basis for the development of specific detection assays for T. equigenitalis. Les lipopolysaccharides (LPSs) provenant de Taylorella equigenitalis, l’agent étiologique de la métrite contagieuse équine, et T. asinigenitalis ont été comparés par électrophorèse sur gel de polyacrylamide avec du sulfate de dodécyl sodique (SDS-PAGE). Les profils de LPSs de 11 souches de T. equigenitalis étaitent similaires, mais différaient des profils de 3 souches de T. asinigenitalis, et les profils de 2 souches de T. asinigenitalis étaient similaires entre eux. Les spécificités sérologiques des LPSs de ces 14 souches ont été examinées par immunobuvardage et immunoessais avec des anticorps monoclonaux (Mabs) contre les LPSs des souches types de T. equigenitalis et T. asinigenitalis et la souche 2329–98 de T. asinigenitalis. Un MAb contre le polysaccharide O (O-PS) du LPS de M. equigenitalis (M2560) a réagit avec les LPSs de toutes les souches de T. equigenitalis mais n’a pas réagit avec les LPSs des 3 souches de T. asinigenitalis ou des 43 bactéries différentes de Taylorella. Trois MAbs dirigés contre le O-PS LPS ou les épitopes du core de la souche type de T. asinigenitalis (M2974, M2982, M3000) ont réagit avec la souche homologue et avec la souche Bd 3751/05 de T. asinigenitalis, mais avec aucune autre bactérie. Cinq MAbs dirigés contre le O-PS du LPS ou les épitopes du core de T. asinigenitalis 2329–98 (M2904, M2907, M2910, M2923, M2929) n’ont réagit qu’avec cette souche. Le spectre des O-PSs des souches types de T. equigenitalis et T. asinigenitalis obtenu par résonnance magnétique nucléaire protonique a fourni des empreintes pour l’identification et la différenciation de ces 2 organismes. Les résultats sérologiques étaient compatibles avec nos résultats précédents qui indiquaient que l’antigène O de la souche type de T. equigenitalis, qui est un polymère linéaire d’unités répétées du disaccharide [→4)-α -L-GulpNAc3NAcA-(1→4)- β-D-ManpNAc3NAcA-(1→], diffère du polymère de l’antigène O de T. asinigenitalis qui est composé d’unités répétées de [3→)- β-D-QuipNAc4NAc-(1→3)- β-D-GlcpNAmA-(1→]. Le O-PS du LPS pourrait être un marqueur spécifique pour l’identification et la différenciation de T. equigenitalis et T. asinigenitalis, et il fournit les éléments de base pour le développement d’épreuves spécifiques de détection de T. equigenitalis. (Traduit par Docteur Serge Messier)
Publication Date: 2010-04-02 PubMed ID: 20357953PubMed Central: PMC2801306
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- Journal Article
- Antibodies
- Contagious Equine Metritis
- Diagnosis
- Diagnostic Technique
- Disease Diagnosis
- Electrophoresis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Equine Diseases
- Equine Health
- Horses
- Immunology
- Infectious Disease
- Laboratory Methods
- Lipopolysaccharide
- Molecular biology
- Monoclonal Antibodies
- Serology
- Spectroscopy
- Veterinary Medicine
- Veterinary Research
Summary
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The research paper discusses the identification and distinction of two bacteria strains, Taylorella equigenitalis and Taylorella asinigenitalis, by analyzing their Lipopolysaccharides (LPSs) structures using serology methods with monoclonal antibodies (MAbs).
Lipopolysaccharides (LPSs) Comparison
- The research used sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) to compare the LPSs from Taylorella equigenitalis and T. asinigenitalis.
- The LPS profile findings, which represents the composition and structure of the bacterial cell wall, showed that 11 strains of T. equigenitalis were similar to each other but different from 3 strains of T. asinigenitalis, which were also similar to each other.
Use of Monoclonal Antibodies in Identification
- The specificities of the LPSs were examined through immunoblotting and enzyme-linked immunosorbent assay utilizing monoclonal antibodies (MAbs), which are identical antibodies made by identical immune cells that were cloned from a unique parent cell.
- When these monoclonal antibodies were exposed to T. equigenitalis, they reacted with the LPS from all T. equigenitalis strains but not with the LPSs from the 3 T. asinigenitalis strains or any other non-Taylorella bacteria.
- Contrarily, three MAbs reacting with T. asinigenitalis only positively identified the same strain and one other, but none of the other bacteria.
Proton Nuclear Magnetic Resonance (NMR) Spectra
- This method was used to provide a ‘fingerprint’ identification by distinguishing between the type strains of T. equigenitalis and T. asinigenitalis.
- The NMR spectra of the O-Polysaccharides (O-PS) of both types of bacteria were fundamentally different, hence aiding in their identification.
Implication of Findings
- The consistency of the serological results with the previously known O-antigen structure of T. equigenitalis bacteria, as well as the distinct structure of T. asinigenitalis, lends support to their findings.
- The research concludes that the LPS O-PS could potentially serve as a specific marker for the identification and differentiation of the two studied Taylorella strains.
- This study could set the stage for the development of precise detection assays for T. equigenitalis, important for controlling and preventing the spread of infections caused by these bacteria.
Cite This Article
APA
Brooks BW, Lutze-Wallace CL, Maclean LL, Vinogradov E, Perry MB.
(2010).
Identification and differentiation of Taylorella equigenitalis and Taylorella asinigenitalis by lipopolysaccharide O-antigen serology using monoclonal antibodies.
Can J Vet Res, 74(1), 18-24.
Publication
Researcher Affiliations
- Canadian Food Inspection Agency, Ottawa Laboratory (Fallowfield), Ottawa, Ontario. brian.brooks@inspection.gc.ca
MeSH Terms
- Animals
- Antibodies, Bacterial / immunology
- Antibodies, Monoclonal / immunology
- Biomarkers
- Carbohydrate Sequence
- Epitopes
- Mice
- Nuclear Magnetic Resonance, Biomolecular
- O Antigens / chemistry
- O Antigens / immunology
- Serotyping
- Taylorella / classification
- Taylorella equigenitalis / classification
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