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[儿童尿路感染病原体大肠杆菌分离株的毒力因子、系统发育群特征及ST131克隆的存在情况研究]

[Investigation of Virulence Factors, Phylogenetic Group Features, and the Presence of ST131 Clone in Escherichia coli Isolates, a Urinary Tract Infection Agent in Children].

作者信息

Kılıç Selin Gamze, Öcal Duygu, Tekeli Alper, Dolapçı İştar

机构信息

Ankara University Faculty of Medicine, Department of Medical Microbiology, Ankara, Türkiye.

Ankara University Faculty of Medicine, Cebeci Training and Research Hospital, Central Laboratory, Ankara, Türkiye.

出版信息

Mikrobiyol Bul. 2023 Oct;57(4):535-552. doi: 10.5578/mb.20239944.

Abstract

Urinary tract infection (UTI) caused by Escherichia coli is a significant health issue in children. Today especially E.coli O25b/ST131, defined as a pandemic clone, is a serious public health problem due to its high virulence and antimicrobial resistance rates. In this study, a total of 200 (100 first and 100 recurrent UTI-causing) E.coli isolates from urine samples sent to the Ankara University School of Medicine Cebeci Training and Research Hospital Central Laboratory between January and September 2021 with the preliminary diagnosis of UTI in pediatric patients aged three to 18 years were analyzed for antimicrobial resistance rates, phylogenetic group distributions, virulence factor frequencies and whether they belong to the O25b/ST131 clone. It is aimed in this study that, the obtained data will shed light on new studies for diagnosis, treatment and prophylaxis options that can be developed for more effective UTI management by contributing to the surveillance studies in our country. Antimicrobial susceptibility of E.coli isolates identified by conventional methods was evaluated by Kirby-Bauer disc diffusion method and extended spectrum beta-lactamase (ESBL) production was evaluated by double disc synergy test. Polymerase chain reaction (PCR) was used for the investigation of phylogenetic grouping, the O25b/ST131 clone, virulence genes and the molecular level classification of the isolates detected as uropathogenic E.coli (UPEC). Pulsed-field gel electrophoresis (PFGE) was performed with the isolates collected at different times from the same patient. The highest antimicrobial resistance rates observed were against ampicillin (n= 100, 50%), cefazolin (n= 99, 49.5%), trimethoprim-sulfamethoxazole (n= 55, 27.5%), amoxicillin-clavulanic acid (n= 43, 21.5%) and cefotaxime (n= 43, 21.5%). In recurrent UTI agents, resistance rates were higher for cefotaxime (n= 29, 29%), trimethoprim-sulfamethoxazole (n= 35, 35%) and cefepime (n= 25, 25%) and in O25b/ST131 isolates (n= 67) the rates were higher for amikacin (n= 3, 4.5%), gentamicin (n= 10, 14.9%) and ciprofloxacin (n= 17, 25.4%) when compared to the first UTI agents and non-O25b/ ST131 isolates (p< 0.05). It was found that 29% (n = 58) of the isolates were multidrug resistant (MDR) and 19% (n = 38) produced ESBL.The rate of recurrent UTI agents was found to be higher among ESBL producing isolates and/or MDR isolates (n= 36, 62% and n= 27, 71%, respectively, p< 0.05). It was found that 45.5% (n= 91) of the isolates were in D, 37.5% (n= 75) in B2, 12.5% (n= 25) in A, and 4.5% (n= 9) in B1 phylogenetic groups and isolates belonging to B2 and D phylogenetic groups had higher antibiotic resistance rates and carried more virulence genes (p< 0.05). Of the isolates, 33.5% (n= 67) were found to belong to the O25b/ST131 clone, no significant difference was found between the O25b/ST131 rates among the first and recurrent UTI agents (p> 0.05). It was determined that the isolates most frequently carry virulence genes for adhesion [fimH 97% (n= 194), papA 57% (n= 114), yfcV 49.5% (n= 99)] and iron uptake systems [fyuA 85.5% (n= 171), chuA 78% (n= 156), iutA 73% (n= 146)]. All virulence factors were detected more frequently in isolates belonging to the O25b/ST131 clone (p< 0.05). Of the isolates, 97% (n= 65) belonging to the O25b/ST131 clone and 27.1% (n= 36) not belonging to this clone were defined as UPEC with molecular analysis (p< 0.0001). Thirty-three isolates belonging to 15 patients were evaluated with PFGE, and it was observed that the latter isolate and the first isolate of eight patients (53%) had the same band profile. Focusing on surveillance, diagnostic testing, treatment algorithms, and preventive measures for E.coli and especially for ST131 clone, which is frequently observed as causative agent in childhood UTIs, will help to manage challenging E.coli infections.

摘要

由大肠杆菌引起的尿路感染(UTI)是儿童中一个重要的健康问题。如今,特别是被定义为大流行克隆株的大肠杆菌O25b/ST131,因其高毒力和抗菌耐药率,成为一个严重的公共卫生问题。在本研究中,对2021年1月至9月间送至安卡拉大学医学院切贝奇培训与研究医院中央实验室的尿样中的200株大肠杆菌分离株(100株初次引起UTI的和100株复发性引起UTI的)进行了分析,这些尿样来自3至18岁儿科患者,初步诊断为UTI,分析内容包括抗菌耐药率、系统发育组分布、毒力因子频率以及它们是否属于O25b/ST131克隆株。本研究旨在通过为我国的监测研究做出贡献,所获得的数据将为新的研究提供线索,以便开发出更有效的UTI管理的诊断、治疗和预防方案。通过 Kirby - Bauer 纸片扩散法评估用传统方法鉴定的大肠杆菌分离株的抗菌敏感性,通过双纸片协同试验评估超广谱β-内酰胺酶(ESBL)的产生。聚合酶链反应(PCR)用于系统发育分组、O25b/ST131克隆株、毒力基因以及检测为尿路致病性大肠杆菌(UPEC)的分离株的分子水平分类的研究。对从同一患者在不同时间收集的分离株进行脉冲场凝胶电泳(PFGE)。观察到的最高抗菌耐药率针对氨苄西林(n = 100,50%)、头孢唑林(n = 99,49.5%)、甲氧苄啶 - 磺胺甲恶唑(n = 55,27.5%)、阿莫西林 - 克拉维酸(n = 43,21.5%)和头孢噻肟(n = 43,21.5%)。在复发性UTI病原体中头孢噻肟(n = 29,29%)、甲氧苄啶 - 磺胺甲恶唑(n = 35,35%)和头孢吡肟(n = 25,25%)的耐药率更高,并且在O25b/ST131分离株(n = 67)中,与初次UTI病原体和非O25b/ST131分离株相比,阿米卡星(n = 3,4.5%)、庆大霉素(n = 10,14.9%)和环丙沙星(n = 17,25.4%)的耐药率更高(p < 0.05)。发现29%(n = 58)的分离株为多重耐药(MDR),19%(n = 38)产生ESBL。在产生ESBL的分离株和/或MDR分离株中复发性UTI病原体的比例更高(分别为n = 36,62%和n = 27,71%,p < 0.05)。发现45.5%(n = 91)的分离株属于D系统发育组,37.5%(n = 75)属于B2,12.5%(n = 25)属于A,4.5%(n = 9)属于B1系统发育组,属于B2和D系统发育组的分离株具有更高的抗生素耐药率且携带更多毒力基因(p < 0.05)。在分离株中,33.5%(n = 67)被发现属于O25b/ST131克隆株,初次和复发性UTI病原体中的O25b/ST131比例之间未发现显著差异(p > 0.05)。确定分离株最常携带的毒力基因是用于黏附的[fimH 97%(n = 194),papA 57%(n = 114),yfcV 49.5%(n = 99)]和用于铁摄取系统的[fyuA 85.5%(n = 171),chuA 78%(n = 156),iutA 73%(n = 146)]。所有毒力因子在属于O25b/ST131克隆株的分离株中检测频率更高(p < 0.05)。通过分子分析,属于O25b/ST131克隆株的分离株中有97%(n = 65)被定义为UPEC,不属于该克隆株的分离株中有27.1%(n = 36)被定义为UPEC(p < 0.0001)。对属于15名患者的33株分离株进行了PFGE评估,并观察到8名患者(53%)的后一株分离株和第一株分离株具有相同的条带图谱。关注大肠杆菌尤其是在儿童UTI中经常作为病原体出现的ST131克隆株的监测、诊断测试、治疗算法和预防措施,将有助于管理具有挑战性的大肠杆菌感染。

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