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目的建立一种能够对由致病性大肠杆菌引发的宿主细胞黏附及擦拭性损伤(A/E损伤)进行半定量分析的方法,为进一步研究其致病性提供评价手段。方法以大肠杆菌黏附HeLa细胞作为感染模型。采用姬姆萨染色和荧光肌动蛋白染色实验,通过菌落计数及肌动蛋白聚集数量定量分析,统计学处理,比较EPEC 43095、EHEC O157∶H7 Sakai株和弱毒株EHEC O157∶H7 00B015以及E.coliDH5α对HeLa细胞的黏附能力,评价它们对细胞的损伤程度。结果EPEC对细胞的黏附及擦拭性损伤程度大于EHEC(P<0.05)。EHEC Sakai和00B015在黏附能力上差异不大(P>0.05),但是,Sakai在肌动蛋白聚集形成能力方面却明显强于00B015(P<0.05)。结论成功建立了一种能够对EPEC或EHEC所引发的宿主细胞黏附及擦拭性损伤进行半定量分析的方法。
Objective To establish a semi-quantitative method capable of semi-quantitative analysis of host cell adhesion and wiping damage (A / E injury) caused by pathogenic Escherichia coli and provide an evaluation method for further study of its pathogenicity. Methods E. coli HeLa cells were used as the model of infection. Using Giemsa staining and fluorescent actin staining experiments, the number of colony counts and actin aggregation quantitative analysis, statistical analysis, comparison EPEC 43095, EHEC O157: H7 Sakai strains and attenuated strains EHEC O157: H700B015 and E. coli DH5α on HeLa cell adhesion capacity, evaluate their damage to cells. Results The adhesion and wiping of EPEC were more severe than that of EHEC (P <0.05). EHEC Sakai and 00B015 showed no significant differences in adhesion (P> 0.05). However, Sakai was significantly stronger than that of 00B015 in actin aggregation (P <0.05). Conclusion A semi-quantitative method for semi-quantitative analysis of host cell adhesion and wiping damage induced by EPEC or EHEC was successfully established.