1Department of Microbiology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran.
2Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran.
Acta Microbiol Immunol Hung. 2023 Jun 21;70(3):206-212. doi: 10.1556/030.2023.02060. Print 2023 Sep 21.
Globally, the spread of carbapenem-resistant strains has limited treatment options for multidrug-resistant (MDR) Pseudomonas aeruginosa infections. This study aimed to determine the role of point mutations as well as the expression level of the oprD gene in the emergence of imipenem-resistant P. aeruginosa strains isolated from patients referred to Ardabil hospitals. A total of 48 imipenem-resistant clinical isolates of P. aeruginosa collected between June 2019 and January 2022 were used in this study. Detection of the oprD gene and its amino acid alterations was performed using the polymerase chain reaction (PCR) and DNA sequencing techniques. The expression level of the oprD gene in imipenem-resistant strains was determined using the real-time quantitative reverse transcription PCR (RT-PCR) method. All imipenem-resistant P. aeruginosa strains were positive for the oprD gene based on the PCR results, and also five selected isolates indicated one or more amino acid alterations. Detected amino acid alterations in the OprD porin were Ala210Ile, Gln202Glu, Ala189Val, Ala186Pro, Leu170Phe, Leu127Val, Thr115Lys, and Ser103Thr. Based on the RT-PCR results, the oprD gene was downregulated in 79.1% of imipenem-resistant P. aeruginosa strains. However, 20.9% of strains showed overexpression of the oprD gene. Probably, resistance to imipenem in these strains is associated with the presence of carbapenemases, AmpC cephalosporinase, or efflux pumps. Owing to the high prevalence of imipenem-resistant P. aeruginosa strains due to various resistance mechanisms in Ardabil hospitals, the implementation of surveillance programs to reduce the spread of these resistant microorganisms along with rational selection and prescription of antibiotics is recommended.
全球范围内,碳青霉烯类耐药菌株的传播使得多重耐药(MDR)铜绿假单胞菌感染的治疗选择有限。本研究旨在确定点突变以及 oprD 基因表达水平在从 Ardabil 医院就诊患者中分离的耐亚胺培南铜绿假单胞菌菌株出现中的作用。本研究共使用了 2019 年 6 月至 2022 年 1 月期间收集的 48 株耐亚胺培南临床分离的铜绿假单胞菌。使用聚合酶链反应(PCR)和 DNA 测序技术检测 oprD 基因及其氨基酸改变。使用实时定量逆转录 PCR(RT-PCR)方法确定耐亚胺培南菌株中 oprD 基因的表达水平。所有耐亚胺培南铜绿假单胞菌菌株的 PCR 结果均为 oprD 基因阳性,并且还在 5 个选定的分离株中检测到一个或多个氨基酸改变。在 OprD 孔蛋白中检测到的氨基酸改变为 Ala210Ile、Gln202Glu、Ala189Val、Ala186Pro、Leu170Phe、Leu127Val、Thr115Lys 和 Ser103Thr。根据 RT-PCR 结果,79.1%的耐亚胺培南铜绿假单胞菌菌株 oprD 基因下调。然而,20.9%的菌株 oprD 基因过表达。这些菌株对亚胺培南的耐药性可能与碳青霉烯酶、AmpC 头孢菌素酶或外排泵的存在有关。由于 Ardabil 医院存在多种耐药机制,导致耐亚胺培南铜绿假单胞菌菌株的高流行率,建议实施监测计划以减少这些耐药微生物的传播,并合理选择和处方抗生素。