从尿液样本中分离出具有多重耐药性的尿路致病性大肠杆菌并对其进行分子鉴定:对尿路感染管理的启示

Bernaitis L , Bharathi Priya , Ezhilarasu A , Revathi P Shenoy
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摘要

由尿路致病性大肠杆菌(UPEC)引起的尿路感染(UTI)给全球健康造成了巨大负担。约 90% 的尿路感染是由 UPEC 引起的,因此了解其致病机制和抗生素耐药性对于制定有效的治疗策略至关重要。本研究旨在调查尿路感染中多重耐药(MDR)大肠杆菌菌株的流行情况,并阐明其耐药性的分子机制。在 2022 年 11 月至 2023 年 2 月的四个月期间,从班加罗尔 Kalyan Nagar 医生诊断中心采集的尿路感染患者标本中共分离出 150 株大肠杆菌。使用伊红亚甲基蓝琼脂和营养琼脂对致病性大肠杆菌进行了分离和筛选,然后通过革兰氏染色和生化特性测试进行鉴定。抗生素敏感性测试使用穆勒-欣顿琼脂和各种抗生素盘进行。对扩谱β-内酰胺酶(ESBL)产生者进行了表型和分子鉴定,并使用聚合酶链式反应(PCR)筛选了特定的抗生素耐药基因。在 150 个大肠杆菌分离物中,发现 57.3% 具有多重耐药性,其中来自住院患者的 35 个分离物均表现出多重耐药性特征。表型和分子分析表明,分离物中存在产 ESBL 的菌株,在 PCR 检测中发现了 blaCTX-M、blaTEM 和 blaSHV 基因。质粒提取和 PCR 检测进一步确定了质粒编码的碳青霉烯酶基因(blaOXA-48、blaNDM-1 和 blaKPC-2)以及其他抗生素耐药基因的存在,包括对氟喹诺酮类药物产生耐药性的基因。研究结果表明,MDR 大肠杆菌菌株在尿路感染中的流行程度令人担忧,尤其是在住院病人中。分子特征描述揭示了导致抗生素耐药性的多种基因机制,包括 ESBLs 和质粒编码耐药基因的存在。这些发现强调,迫切需要采取监控和感染控制措施来减少 MDR 病原体的传播,并开发替代治疗策略来对抗尿路感染中的抗生素耐药性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Isolation and molecular characterization of multi-drug resistant uropathogenic Escherichia coli from urine samples: Insights into urinary tract infection management
Urinary tract infections (UTIs) caused by uropathogenic Escherichia coli (UPEC) present a significant global health burden. With UPEC responsible for approximately 90 % of UTIs, understanding its mechanisms of virulence and antibiotic resistance is paramount for effective treatment strategies. This study aimed to investigate the prevalence of multi-drug resistant (MDR) E. coli strains in urinary tract infections and to elucidate the molecular mechanisms underlying their resistance. A total of 150 E. coli isolates were obtained from urinary tract infection specimens collected patients at Doctor’s Diagnostic Centre in Kalyan Nagar, Bangalore over four months November 2022 to February 2023. Isolation and screening of pathogenic E. coli were conducted using Eosin Methylene Blue Agar and Nutrient Agar, followed by identification through gram staining and biochemical characterization tests. Antibiotic susceptibility testing was performed using Mueller-Hinton agar and various antibiotic discs. Phenotypic and molecular identification of extended-spectrum beta-lactamase (ESBL) producers were conducted, along with the screening for specific antibiotic resistance genes using polymerase chain reaction (PCR). Of the 150 E. coli isolates, 57.3 % were found to be multi-drug resistant, with all 35 isolates from hospitalized patients exhibiting MDR characteristics. Phenotypic and molecular analyses revealed the presence of ESBL-producing isolates, with the blaCTX-M, blaTEM, and blaSHV genes detected in PCR assays. Plasmid extraction and PCR assays further identified the presence of plasmid-encoded carbapenemase genes (blaOXA-48, blaNDM-1, blaKPC-2), as well as other antibiotic resistance genes including those conferring resistance to fluoroquinolones. The results underscore the alarming prevalence of MDR E. coli strains in urinary tract infections, particularly in hospitalized patients. Molecular characterization revealed the diverse genetic mechanisms contributing to antibiotic resistance, including the presence of ESBLs and plasmid-encoded resistance genes. These findings emphasize the urgent need for surveillance and infection control measures to mitigate the spread of MDR pathogens and the development of alternative therapeutic strategies to combat antibiotic resistance in UTIs.
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