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1. 第三军医大学医学检验系临床微生物学及免疫学教研室
2. 江苏省疾病预防控制中心
纸质出版日期:2009,
网络出版日期:2009-04-30,
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王海光, 顾江, 余抒, 等. 致病性大肠杆菌引发宿主细胞A/E损伤分析方法的建立[J]. 陆军军医大学学报, 2009,31(8):659-662.
WANG HAI-GUANG, GU JIANG, YU SHU, et al. Evaluation of A/E lesions in HeLa cells caused by pathogenic E.coli: Giemsa staining and fluorescence actin staining. [J]. Journal of army medical university, 2009, 31(8): 659-662.
王海光, 顾江, 余抒, 等. 致病性大肠杆菌引发宿主细胞A/E损伤分析方法的建立[J]. 陆军军医大学学报, 2009,31(8):659-662. DOI:
WANG HAI-GUANG, GU JIANG, YU SHU, et al. Evaluation of A/E lesions in HeLa cells caused by pathogenic E.coli: Giemsa staining and fluorescence actin staining. [J]. Journal of army medical university, 2009, 31(8): 659-662. DOI:
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.coli
DH5α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 to analyze the attaching and effacing (A/E) lesions on host cells caused by pathogenic
Escherichia coli
(
E. coli
). Methods HeLa cells were infected with enteropathogenic or enterohemorrhagic
E. coli
strains (EPEC 43095
EHEC O157∶H7 Sakai
EHEC O157∶H7 00B015 and
E. coli
DH5α) and visualized by Giemsa staining and fluorescence actin staining (FAS). Then the numbers of microcolonies and actin polymerizations formed on the cells were counted and calculated. Results All
EPEC induced stronger A/E lesions than EHEC on HeLa cells (
P
<
0.05). The EPEC 43095 caused the strongest A/E lesions among these
E. coli
strains detected. Although the ability of EHEC Sakai to attach to HeLa cells was similar to that of EHEC 00B015 (P>0.05)
EHEC Sakai induced stronger actin polymerizations in host cells than EHEC 00B015 (
P
<
0.05). Conclusion A semi-quantitative method to analyze the A/E lesions of host cells caused by pathogenic
E. coli
is successfully established. This work contributes to further studies on pathogenicity of the pathogenic
E.coli
.
肠致病性大肠杆菌肠出血性大肠杆菌黏附和擦拭性损伤
enteropathogenic Escherichia colienterohemorrhagic Escherichia coliattaching and effacing lesions
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