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基于 PCR 的 RFLP 和 ERIC-PCR 图谱显示与埃及多药耐药相关的幽门螺杆菌菌株。

PCR-based RFLP and ERIC-PCR patterns of Helicobacter pylori strains linked to multidrug resistance in Egypt.

机构信息

Microbiology and Immunology Department, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt.

Department of Biochemistry and Nutrition, Faculty of Women for Arts, Sciences and Education, Ain Shams University, Heliopolis, Cairo, Egypt.

出版信息

Sci Rep. 2024 Sep 27;14(1):22273. doi: 10.1038/s41598-024-72289-z.

Abstract

H. pylori infects approximately 50% of the world's population that causes chronic gastritis, and may lead to peptic ulcer disease (PUD). H. pylori-induced chronic infections are associated with gastric adenocarcinoma and low-grade gastric lymphoma. In Egypt, H. pylori strains are widespread and became resistant to antimicrobial agents, thus advanced typing methods are needed to differentiate infectious strains that are resistant to antibiotics, and therefore earlier prognosis and infection control. The main objectives were (i) to determine susceptibility of infectious H. pylori strains to some antimicrobial agents that are currently used in eradication therapy in Egypt; (ii) to identify diverse strains commonly detected in the gastrointestinal (GIT) endoscopy units in Egypt through phenotypic and genotypic analyses. In this observational study we isolated 167 isolates from 232 gastric biopsies (antrum and corpus) of patients who were admitted to the upper GIT endoscopy units in five governmental Egyptian hospitals. Antimicrobial susceptibility patterns were investigated using Kirby Bauer disc diffusion and agar dilution Minimum Inhibitory Concentrations (MICs) methods. Phenotypic characterization was based on biotyping and antibiogram typing techniques. Genotypic characterization was carried out using PCR-based Restriction Fragment Length Polymorphism (RFLP) and Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR analyses. H. pylori isolates were highly resistant to diverse antimicrobial agents including Metronidazole, Fluoroquinolones, Macrolides, Amoxycillin, Tetracycline and Gentamicin. Two factors contributed to the increased resistance of H. pylori to the conventional therapy seen in Egypt: (i) Metronidazole and Amoxycillin are inexpensive and available drugs being abused by patients; (ii) the regional prescribing practice of Macrolids commonly used to treat upper respiratory and urinary tract infections. Five different biotypes were identified depending on the ability of the isolates to synthesize different enzymes. Nine antibiogram types were identified. PCR-RFLP analysis revealed fifteen different fingerprints while ERIC-PCR revealed 22 fingerprints. Biotyping alone or in combination with antibiogram typing are highly useful molecular tools in the prognosis of strain relatedness. PCR-RFLP and ERIC-PCR acquired good discriminatory power for identifying H. pylori infectious sub-types.

摘要

幽门螺杆菌感染了世界上约 50%的人口,导致慢性胃炎,并可能导致消化性溃疡病(PUD)。幽门螺杆菌引起的慢性感染与胃腺癌和低级别胃淋巴瘤有关。在埃及,幽门螺杆菌菌株广泛存在,并对抗菌药物产生了耐药性,因此需要先进的分型方法来区分对抗生素耐药的传染性菌株,从而更早地进行预后判断和感染控制。主要目标是:(i)确定目前在埃及根除治疗中使用的一些抗菌药物对感染性幽门螺杆菌菌株的敏感性;(ii)通过表型和基因型分析,鉴定在埃及胃肠道内窥镜单位中常见的多种菌株。在这项观察性研究中,我们从 5 家埃及政府医院的上胃肠道内窥镜单位收治的 232 例胃活检(胃窦和胃体)中分离出 167 株幽门螺杆菌。采用 Kirby Bauer 纸片扩散法和琼脂稀释最低抑菌浓度(MIC)法检测抗菌药物敏感性模式。表型特征基于生物分型和抗生素药敏谱分型技术。基因型特征采用基于 PCR 的限制性片段长度多态性(RFLP)和肠杆菌重复基因间一致性(ERIC)-PCR 分析进行。幽门螺杆菌分离株对多种抗菌药物高度耐药,包括甲硝唑、氟喹诺酮类、大环内酯类、阿莫西林、四环素和庆大霉素。有两个因素导致埃及幽门螺杆菌对传统治疗的耐药性增加:(i)甲硝唑和阿莫西林价格低廉且易于获得,因此被患者滥用;(ii)大环内酯类药物是治疗上呼吸道和尿路感染的常用药物,导致该区域的处方习惯。根据分离株合成不同酶的能力,确定了 5 种不同的生物型。鉴定了 9 种抗生素药敏谱类型。PCR-RFLP 分析显示了 15 种不同的指纹图谱,而 ERIC-PCR 显示了 22 种指纹图谱。生物分型单独或与抗生素药敏谱分型相结合,是预测菌株相关性的高度有用的分子工具。PCR-RFLP 和 ERIC-PCR 对鉴定幽门螺杆菌感染亚型具有良好的鉴别力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e7ed/11436738/ea5179e3f18e/41598_2024_72289_Fig1_HTML.jpg

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