Detection of Autotransporter Genes and Evaluation of the Relationship Between the Autotransporters and Biofilm Formation in Uropathogenic Escherichia Coli Isolated From Urinary Tract Infections

M. A. Asadi Karam, M. Habibi
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Abstract

Background: Uropathogenic Escherichia coli (UPEC) are among the pathogens causing urinary tract infections (UTIs). UPEC strains utilize autotransporter (AT) proteins for pathogenesis, especially the development of biofilm in the urinary tract. Objective: There is no general data about the frequency of ATs among the UPECs, thus we decided to assess the frequency and AT patterns in patients with UPEC recovered from UTI, as well as to find the possible correlation between the ATs and biofilm. Materials and Methods: Totally, 200 Escherichia coli were recovered from the urine of outpatients and inpatients with UTIs in Iran. Biofilm production was assessed by a phenotypic method. Extraction of DNA was done using the boiling method, and the extracted products were used for polymerase chain reaction (PCR). Amplification of AT genes was done by PCR with an optimized program in a thermal cycler apparatus. Then, the amplified genes were evaluated by electrophoresis. Results: Among the UPEC isolates, 63.5% and 36.5% were isolated from cystitis and pyelonephritis patients, respectively. In addition, the majority of UPECs (66.5%) were associated with hospitalized patients. Totally, 92% of isolates produced biofilm, of which 53.5%, 21.5%, and 17% were weak, moderate, and strong biofilm producers, respectively. Among investigated ATs, the most frequent genes were upaH (89%), upaB (87%), upaG (79%), vat (78%), sat (69%), upaC (67%), and ag43 (37%). There were 37 AT patterns among UPECs, and pattern 1 with the presence of all AT genes was the most common. It was found that the prevalence of upaB and upaG was significantly higher among the cystitis isolates compared to pyelonephritis isolates. Further, no significant difference was observed between the AT genes, except for upaC (P=0.003), and the intensity of biofilm formation in UPEC isolates. Conclusion: According to the high prevalence of AT genes among UPECs isolated from inpatients and outpatients, these factors can be ideal vaccine candidates for designing effective vaccines against the isolates. Evaluation of the efficacy of these ATs against UPECs is under investigation.
尿路感染尿路致病性大肠杆菌中自转运蛋白基因的检测及其与生物膜形成的关系
背景:尿路致病性大肠杆菌(UPEC)是引起尿路感染(uti)的病原体之一。UPEC菌株利用自身转运蛋白(AT)致病,特别是在尿路中形成生物膜。目的:UPEC患者中AT的发生频率没有一般的数据,因此我们决定评估UPEC患者UTI恢复后AT的发生频率和AT的模式,并寻找AT与生物膜之间可能的相关性。材料与方法:从伊朗门诊和住院尿路感染患者尿液中共检出大肠杆菌200株。生物膜的生成通过表型方法进行评估。采用煮沸法提取DNA,提取产物用于聚合酶链反应(PCR)。AT基因在热循环装置中用优化程序进行PCR扩增。然后,对扩增的基因进行电泳鉴定。结果:UPEC分离株中,膀胱炎和肾盂肾炎患者分离株分别占63.5%和36.5%。此外,大多数UPECs(66.5%)与住院患者相关。92%的分离菌产生生物膜,其中53.5%为弱生膜菌,21.5%为中等生膜菌,17%为强生膜菌。在研究的ATs中,最常见的基因是upaH(89%)、upaB(87%)、upaG(79%)、vat(78%)、sat(69%)、upaC(67%)和ag43(37%)。upec共有37种AT模式,其中AT基因全部存在的模式1最为常见。结果发现,与肾盂肾炎分离株相比,膀胱炎分离株中upaB和upaG的患病率明显较高。此外,除了upaC (P=0.003)外,AT基因与UPEC分离株的生物膜形成强度之间没有显著差异。结论:鉴于AT基因在门诊和住院upec中较高的流行率,这些因素可作为设计有效疫苗的理想候选疫苗。目前正在对这些ATs对upec的疗效进行评估。
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