尿路致病性大肠埃希菌外膜蛋白T敲除株的构建及功能评价  被引量:5

Construction and functional studies of uropathogenic E.coli strains with ompT gene knockout

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作  者:赵铁[1] 方幸幸[1] 刘晓露[1] 彭亮[1] 龙敏[1] 张文炳[1] 罗军[1] 曹虹[1] 

机构地区:[1]南方医科大学公共卫生与热带医学学院医学微生物系,广东广州510515

出  处:《南方医科大学学报》2012年第7期956-959,共4页Journal of Southern Medical University

基  金:国家自然科学基金(30972637);南方医科大学公卫学院院长基金(GW201101)

摘  要:目的探索外膜蛋白T(OmpT)基因在尿路致病性大肠埃希菌(UPEC)CFT073致尿路感染中的作用。方法利用RED重组技术构建UPEC CFT073 ompT基因缺失株(命名为COTD),并建立小鼠急性尿路感染模型,体内外实验比较野生株与敲除株之间在膀胱组织的定植能力。结果 PCR分析及测序鉴定证实ompT基因缺失株构建成功;体外实验结果显示野生株粘附率为(8.3±1.9)%,敲除株粘附率为(6.7±2.2)%,敲除株粘附率明显低于野生株(P<0.05)。体内实验膀胱组织内,野生株定植细菌数为(7±2)×105cfu,敲除株定植细菌数为(17±8)×104cfu,ompT敲除株定植于膀胱组织的能力明显降低(P<0.05)。结论证实OmpT作为毒力因子参与细菌定植膀胱组织,在UPEC致尿路感染的过程中发挥重要作用。Objective To explore the role of ompT gene in uropathogenic E. coli (UPEC) CFT073 strain in urinary tract infection (UTI). Methods An ompT deletion mutant (COTD) was generated by A Red recombineering in the UPEC CFT073 strain, which was characterized by PCR and sequencing. C57B/L6 mouse models of acute UTI with the mutant and wild-type strains were established to compare the colonization abilities of the two strains in the bladder. The adhesion of CFT073 mutant to human unthelial 5637 cells was also investigated in vitro. Results PCR and DNA sequencing confirmed the loss of ompT gene in the mutant COTD. The in vitro adhesion rate of the mutant strain COTD to 5637 cells was (6.7±2.2)%, significantly lower than that of (8.3±1.9)% of the wild-type strain (P〈0.05). In the murine models of acute UTL the mutant strain showed a mean colonization number of about (17±8)×10^4 cfu, which was significantly lower than that of (7±2)×10^5 cfu of the wide-type CFT073 strain (P〈0.05). Conclusion OmpT gene can be involved in the colonization of UPEC in the bladder tissue and plays an important role in the pathogenesis of UPEC-induced UTI.

关 键 词:尿路致病性大肠埃希菌 外膜蛋白T 尿路感染 定植 

分 类 号:R378[医药卫生—病原生物学]

 

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