HEp-2细胞的粘附特征及集聚性粘附相关基因分析.docVIP

我国EAEC分离株对HEp-2细胞的粘附特征及集聚性粘附相关基因分析 刘学通，陈强，孙晖，赵爱兰，徐建国，熊衍文 【摘要】：目的 了解我国EAEC分离株的HEp-2细胞粘附特征、毒力基因和集聚性粘附基因情况，探讨建立有效检测方法的可能。 方法 选择31株分离自腹泻患者粪便的大肠杆菌，采用HEp-2细胞粘附试验，获得菌株的粘附表型；采用PCR方法检测毒力基因（astA、aggR、pic）及集聚性粘附相关基因（aggA、aafA、agg3A、hdaA）。 结果不同菌株在HEp-2细胞粘附试验中表现为不同的粘附表型，所含有的毒力基因及粘附相关基因不同。集聚性粘附（AA）表型。 结论 HEp-2细胞粘附试验仍然是检测EAEC的金标准；aggR基因仍不失为一种较好的PCR诊断靶基因。 【关键词】 肠集聚性大肠杆菌；粘附表型；粘附基因；HEp-2细胞 Adherence Characteristics to HEp-2 Cells and Aggregative Adherence Associated Genes of EAEC Strains Isolated from China Liu Xue-tong, Chen Qiang, Sun Hui, Zhao Ai-lan, Xu Jian-guo, Xiong Yan-wen 【ABSTRACT】 Objective To understand adherence and virulence gene characteristics of EAEC isolates to explore the possibility of establishing PCR detection methods. Methods 31 Escherichia coli strains isolated from diarrhea patients’ stools were selected.The adherence phenotypes were tested by HEp-2 cells adherence assay. The virulence genes(astA、aggR、pic) and aggregative adherence associated genes (aggA、aafA、agg3A、hdaA) were tested by PCR. Results Different strains showed different adherence phenotype to HEp-2 cells and different virulence genes and aggregative adherence associated genesprofiles. Thirteen of the tested strains are aggR positive, seven among which showed aggregative adherence (AA) phenotype. Five of the seven strains can detect aggregative adherence associated genes. Besides, we found an atypical EAEC strain which was aggR negative but showed AA phenotype to HEp-2 cells. In addition, there were also strains which had the aggregative adherence associated gene hdaA but didn’t adhere to HEp-2 cells. Conclusions EAEC is a highly heterogeneous group of E. coli.; HEp-2 cells adherence assay is still the ‘gold standard’ to identify EAEC; the aggR gene is still a good target gene for PCR diagnostic. To establish a multiplex PCR which combines other genes on the basis of aggR for EAEC detection has important practical significance. 【KEY WORDS】 EAEC; adherence phenotype; adherence a