使用单克隆抗体通过脂多糖O抗原血清学鉴定和区分马生殖道泰勒菌和驴生殖道泰勒菌。
Identification and differentiation of Taylorella equigenitalis and Taylorella asinigenitalis by lipopolysaccharide O-antigen serology using monoclonal antibodies.
作者信息
Brooks Brian W, Lutze-Wallace Cheryl L, Maclean Leann L, Vinogradov Evgeny, Perry Malcolm B
机构信息
Canadian Food Inspection Agency, Ottawa Laboratory (Fallowfield), Ottawa, Ontario.
出版信息
Can J Vet Res. 2010 Jan;74(1):18-24.
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.
通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）对马生殖道支原体（传染性马子宫内膜炎的病原体）和猪生殖道支原体的脂多糖（LPS）进行了比较。11株马生殖道支原体菌株的脂多糖谱相似，但与3株猪生殖道支原体菌株的谱不同，且2株猪生殖道支原体菌株的谱彼此相似。用针对马生殖道支原体和猪生殖道支原体模式菌株以及猪生殖道支原体菌株2329-98的LPS的单克隆抗体（MAb），通过免疫印迹和酶联免疫吸附测定法检测了这14株菌株LPS的血清学特异性。一种针对马生殖道支原体LPS O-多糖（O-PS）的单克隆抗体（M2560）与所有马生殖道支原体菌株的LPS发生反应，但不与3株猪生殖道支原体菌株的LPS或43株非泰勒菌属细菌的LPS发生反应。三种针对猪生殖道支原体模式菌株LPS O-PS或核心表位的单克隆抗体（M2974、M2982、M3000）与同源菌株和猪生殖道支原体菌株Bd 3751/05发生反应，但不与任何其他细菌发生反应。五种针对猪生殖道支原体2329-98 LPS O-PS或核心表位的单克隆抗体（M2904、M2907、M2910、M2923、M2929）仅与该菌株发生反应。马生殖道支原体和猪生殖道支原体模式菌株O-PS的质子核磁共振谱提供了这两种生物体的指纹识别和区分。血清学结果与我们之前的发现一致，即马生殖道支原体模式菌株的O抗原是二糖重复单元[-->4)-α-L-古洛糖胺3NAcA-(1-->4)-β-D-甘露糖胺3NAcA-(1-->]的线性聚合物，与由重复单元[-->3)-β-D-喹诺糖胺4NAc-(1-->3)-β-D-葡萄糖胺A-(1-->]组成的猪生殖道支原体O抗原聚合物不同。脂多糖O-PS可能是鉴定和区分马生殖道支原体和猪生殖道支原体的特异性标志物，并为开发马生殖道支原体特异性检测方法提供依据。
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