Phenotypic and molecular characterization of extended spectrum- and metallo- beta lactamase producing Pseudomonas aeruginosa clinical isolates from Egypt.

IF 5.4 2区 医学 Q1 INFECTIOUS DISEASES Infection Pub Date : 2024-12-01 Epub Date: 2024-06-02 DOI:10.1007/s15010-024-02297-8
Eva A Edward, Marwa R El Shehawy, Alaa Abouelfetouh, Elsayed Aboulmagd
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Abstract

Background: Antimicrobial resistance among Pseudomonas aeruginosa (P. aeruginosa), a leading cause of nosocomial infections worldwide, is escalating. This study investigated the prevalence of extended-spectrum β-lactamases (ESBLs) and metallo-β-lactamases (MBLs) among 104 P. aeruginosa clinical isolates from Alexandria Main University Hospital, Alexandria, Egypt.

Methods: Antimicrobial susceptibility testing was performed using agar dilution technique, or broth microdilution method in case of colistin. ESBL and MBL prevalence was assessed phenotypically and genotypically using polymerase chain reaction (PCR). The role of plasmids in mediating resistance to extended-spectrum β-lactams was studied via transformation technique using plasmids isolated from ceftazidime-resistant isolates.

Results: Antimicrobial susceptibility testing revealed alarming resistance rates to carbapenems, cephalosporins, and fluoroquinolones. Using PCR as the gold standard, phenotypic methods underestimated ESBL production while overestimating MBL production. Eighty-five isolates (81.7%) possessed only ESBL encoding genes, among which 69 isolates harbored a single ESBL gene [blaOXA-10 (n = 67) and blaPER (n = 2)]. Four ESBL-genotype combinations were detected: blaPER + blaOXA-10 (n = 8), blaVEB-1 + blaOXA-10 (n = 6), blaPSE + blaOXA-10 (n = 1), and blaPER + blaVEB-1 + blaOXA-10 (n = 1). Three isolates (2.9%) possessed only the MBL encoding gene blaVIM. Three ESBL + MBL- genotype combinations: blaOXA-10 + blaAIM, blaOXA-10 + blaVIM, and blaPER + blaOXA-10 + blaAIM were detected in 2, 1 and 1 isolate(s), respectively. Five plasmid preparations harboring blaVEB-1 and blaOXA-10 were successfully transformed into chemically competent Escherichia coli DH5α with transformation efficiencies ranging between 6.8 × 10 3 and 3.7 × 10 4    CFU/μg DNA plasmid. Selected tested transformants were ceftazidime-resistant and harbored plasmids carrying blaOXA-10.

Conclusions: The study highlights the importance of the expeditious characterization of ESBLs and MBLs using genotypic methods among P. aeruginosa clinical isolates to hinder the development and dissemination of multidrug resistant strains.

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埃及铜绿假单胞菌临床分离株产广谱和金属β内酰胺酶的表型和分子特征。
背景:铜绿假单胞菌(P. aeruginosa)是全球范围内导致医院内感染的主要病因,其抗菌药耐药性正在不断升级。本研究调查了埃及亚历山大市亚历山大主大学医院 104 例铜绿假单胞菌临床分离物中扩展谱β-内酰胺酶(ESBLs)和金属β-内酰胺酶(MBLs)的流行情况:方法:使用琼脂稀释技术进行抗菌药物敏感性测试,如果使用的是可乐定,则使用肉汤微量稀释法。使用聚合酶链反应(PCR)对 ESBL 和 MBL 的流行率进行表型和基因型评估。利用从头孢他啶耐药分离物中分离的质粒,通过转化技术研究了质粒在介导对广谱β-内酰胺类药物耐药性中的作用:结果:抗菌药物药敏试验显示,对碳青霉烯类、头孢菌素类和氟喹诺酮类药物的耐药率令人担忧。采用 PCR 作为金标准,表型方法低估了 ESBL 的产生,同时高估了 MBL 的产生。85 个分离株(81.7%)仅拥有 ESBL 编码基因,其中 69 个分离株携带单一 ESBL 基因[blaOXA-10(n = 67)和 blaPER(n = 2)]。检测到四种 ESBL 基因型组合:blaPER + blaOXA-10(n = 8)、blaVEB-1 + blaOXA-10(n = 6)、blaPSE + blaOXA-10(n = 1)和 blaPER + blaVEB-1 + blaOXA-10(n = 1)。三个分离物(2.9%)只拥有 MBL 编码基因 blaVIM。在 2 个、1 个和 1 个分离物中分别检测到三种 ESBL + MBL 基因型组合:blaOXA-10 + blaAIM、blaOXA-10 + blaVIM 和 blaPER + blaOXA-10 + blaAIM。含 blaVEB-1 和 blaOXA-10 的五种质粒制备物被成功转化到具有化学能力的大肠杆菌 DH5α 中,转化效率介于 6.8 × 10 3 和 3.7 × 10 4 CFU/μg DNA 质粒之间。经过测试的转化子具有头孢他啶抗性,并携带 blaOXA-10 质粒:本研究强调了在铜绿假单胞菌临床分离株中使用基因分型方法快速鉴定 ESBLs 和 MBLs 的重要性,以阻止耐多药菌株的发展和传播。
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来源期刊
Infection
Infection 医学-传染病学
CiteScore
12.50
自引率
1.30%
发文量
224
审稿时长
6-12 weeks
期刊介绍: Infection is a journal dedicated to serving as a global forum for the presentation and discussion of clinically relevant information on infectious diseases. Its primary goal is to engage readers and contributors from various regions around the world in the exchange of knowledge about the etiology, pathogenesis, diagnosis, and treatment of infectious diseases, both in outpatient and inpatient settings. The journal covers a wide range of topics, including: Etiology: The study of the causes of infectious diseases. Pathogenesis: The process by which an infectious agent causes disease. Diagnosis: The methods and techniques used to identify infectious diseases. Treatment: The medical interventions and strategies employed to treat infectious diseases. Public Health: Issues of local, regional, or international significance related to infectious diseases, including prevention, control, and management strategies. Hospital Epidemiology: The study of the spread of infectious diseases within healthcare settings and the measures to prevent nosocomial infections. In addition to these, Infection also includes a specialized "Images" section, which focuses on high-quality visual content, such as images, photographs, and microscopic slides, accompanied by brief abstracts. This section is designed to highlight the clinical and diagnostic value of visual aids in the field of infectious diseases, as many conditions present with characteristic clinical signs that can be diagnosed through inspection, and imaging and microscopy are crucial for accurate diagnosis. The journal's comprehensive approach ensures that it remains a valuable resource for healthcare professionals and researchers in the field of infectious diseases.
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