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目的检测伴放线放线杆菌(Aggregatibacter actinomycetemcomitans,Aa)血清型b菌株细胞中是否存在磷酸胆碱。方法采用免疫荧光显微镜技术、狭线印迹法、十二烷硫酸钠-聚丙烯酰胺凝胶电泳和免疫印迹法等方法,用磷酸胆碱特异性单克隆抗体TEPC-15作为检测抗体,检测12株Aa血清型b菌株中的磷酸胆碱。结果通过狭线印迹法在Aa中可以检测到磷酸胆碱,12个血清型b菌株中,5个菌株为磷酸胆碱阳性,占总数的41.7%;免疫荧光显微镜观察结果提示该结构位于Aa的表面,聚丙烯酰胺凝胶电泳免疫印迹法显示,磷酸胆碱附着的结构分子量大小约为9 kDa,免疫荧光显微镜和电泳免疫印迹法的阳性菌与狭线印迹法的阳性菌相同。结论 Aa血清型b菌株细胞中可以检测到磷酸胆碱,该结构可能位于Aa的表面。
Objective To detect the presence or absence of phosphorylcholine in the cells of Aggregatibacter actinomycetemcomitans (Aa) serogroup b. Methods Using immunofluorescence microscopy, slot blotting, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the phosphocholine-specific monoclonal antibody TEPC-15 was used as detection antibody to detect 12 strains Phosphorylcholine in Aa serotype b strains. Results Choline chloride could be detected in Aa by the slot blotting method. Of the 12 serotype b strains, 5 strains were positive for phosphorylcholine, accounting for 41.7% of the total. Immunofluorescence microscopy indicated that the structure was located in Aa Surface and polyacrylamide gel electrophoresis showed that the structure molecular weight of phosphocholine was about 9 kDa. The positive bacteria of immunofluorescence microscopy and electrophoresis immunoblotting were the same as the positive bacteria of slot blot. Conclusions Phosphocholine can be detected in the cells of Aa serotype b strain, which may be located on the surface of Aa.