mexB Efflux Pump Gene Expression, Activity and Biofilm Formation in Clinical Isolates of Pseudomonas Aeruginosa

Официальное название

Relationship Between mexB Efflux Pump Gene Expression, Efflux Pump Activity and Biofilm Formation in Multidrug Resistant Clinical Isolates of Pseudomonas Aeruginosa

Обзор

1. Detection of expression level of mexB efflux pump gene in multidrug resistant clinical isolates of Pseudomonas aeruginosa by real time PCR(QRT-PCR). 2. Phenotypic detection of efflux pump activity in multidrug resistant clinical isolates of Pseudomonas aeruginosa. 3. Phenotypic detection of biofilm formation and its degree (strong-moderate-weak) in multidrug resistant clinical isolates of Pseudomonas aeruginosa. 4. Relationship between expression level of mexB efflux pump gene and degree of biofilm formation in multidrug resistant clinical isolates of Pseudomonas aeruginosa.

Подробное описание

Pseudomonas aeruginosa is an opportunistic gram-negative pathogen, causing life threatening nosocomial infections including respiratory system, urinary system and skin wounds (1). P. aeruginosa exhibits resistance to a range of antibiotic classes, including aminoglycosides, carbapenems, beta-lactams, quinolones, and cephalosporins (2). Frequent occurrence of drug resistance is due to many virulence factors in P. aeruginosa such as flagella, pili, lipopolysaccharides, secreted enzymes like DNase and lipase, toxins, and pigments as pyocyanin which play crucial roles in tissue damage and immune suppression. Multiple resistant mechanisms are developed by P. aeruginosa (3). Two prominent mechanisms, biofilm formation and efflux pump activity among other mechanisms, play an important role in persistence and antibiotic resistance (4). Biofilm formation produced by P. aeruginosa is a complex phenomenon that promotes antibiotic resistance and shields the pathogen from the host immune system. This results in severe clinical outcomes in critically ill patients (5). There are about 12 resistance-nodulation-division (RND) families of efflux pumps in P. aeruginosa. The mexAB-oprM multidrug efflux pump system of P. aeruginosa is involved in resistance (6). Efflux pumps play a role in biofilm formation by influencing Physical-chemical interactions, mobility, gene regulation, quorum sensing (QS), extracellular polymeric substances (EPS), and toxic compound extrusion (6)(7). It has been demonstrated that MexAB-OprM plays a role in the resistance of aztreonam, gentamicin, tetracycline and tobramycin in biofilm structures of P. aeruginosa (8). MexB is the most critical and specific gene for efflux activity within the MexAB-OprM system and contributes to antibiotic resistance in Pseudomonas aeruginosa (9).

Вмешательства

• Генная терапия Real time PCR
  Clinical isolates of Pseudomonas aeruginosa will undergo RNA extraction followed by cDNA (complementary DNA) synthesis. Quantitative real-time PCR will be performed using specific primers for the mexB efflux pump gene, and SYBR Green will be used to quantify gene expression. Housekeeping genes (e.g., rpoD or 16S rRNA) will serve as internal controls. The procedure allows precise quantification of mexB expression levels in each isolate.

Первичные конечные точки

• Detection of mexB efflux pump gene expression level in multidrug resistant clinical isolates of Pseudomonas aeruginosa [Срок оценки: one year]

Вторичные конечные точки (2)

• Detection of efflux pump activity in multidrug resistant clinical isolates of Pseudomonas aeruginosa [Срок оценки: one year]
• Detection of degree of biofilm formation (strong-moderate-weak) in multidrug resistant clinical isolates of Pseudomonas aeruginosa [Срок оценки: one year]

Критерии участия

Критерии включения

• Clinical isolates confirmed as Pseudomonas aeruginosa by microbiological testing.
• Isolates collected from different clinical specimens (e.g., wound swabs, sputum, urine, blood, and catheters).

Критерии исключения

• Bacterial species other than Pseudomonas aeruginosa.
• Repeated clinical isolates from the same patient.
• Repeated clinical specimens from different sources of the same patient.

Критерии приведены из реестра в оригинале (на английском). Окончательную оценку соответствия проводит исследовательский центр.

Здоровые добровольцы: Нет

Дизайн исследования

Модель наблюдения

Когортное

Центры проведения

Список центров уточняется — проверьте первичный протокол.

Публикации

• Li XF, Shi HQ, Liang Y, Li J, Jiang B, Song GB. Interaction of biofilm and efflux pump in clinical isolates of carbapenem resistant P. aeruginosa. Eur Rev Med Pharmacol Sci. 2022 Mar;26(5):1729-1737. doi: 10.26355/eurrev_202203_28242. PMID 35302222
• Soltani Borchaloee A, Moosakazemi Mohammadi LS, Khosh Ravesh R, Allameh SF, Tabatabaie Poya FS, Fatehi MA. Prevalence of Biofilm and Efflux Pump Genes Expression by PCR and Antibiotic Resistance Pattern in Pseudomonas Aeruginosa. Arch Razi Inst. 2024 Dec 31;79(6):1281-1286. doi: 10.32592/ARI.2024.79.6.1281. eCollection 2024 Dec. PMID 40599435
• Liao C, Huang X, Wang Q, Yao D, Lu W. Virulence Factors of Pseudomonas Aeruginosa and Antivirulence Strategies to Combat Its Drug Resistance. Front Cell Infect Microbiol. 2022 Jul 6;12:926758. doi: 10.3389/fcimb.2022.926758. eCollection 2022. PMID 35873152
• Ugwuanyi FC, Ajayi A, Ojo DA, Adeleye AI, Smith SI. Evaluation of efflux pump activity and biofilm formation in multidrug resistant clinical isolates of Pseudomonas aeruginosa isolated from a Federal Medical Center in Nigeria. Ann Clin Microbiol Antimicrob. 2021 Feb 2;20(1):11. doi: 10.1186/s12941-021-00417-y. PMID 33531042
• Karballaei Mirzahosseini H, Hadadi-Fishani M, Morshedi K, Khaledi A. Meta-Analysis of Biofilm Formation, Antibiotic Resistance Pattern, and Biofilm-Related Genes in Pseudomonas aeruginosa Isolated from Clinical Samples. Microb Drug Resist. 2020 Jul;26(7):815-824. doi: 10.1089/mdr.2019.0274. Epub 2020 Jan 23. PMID 31976811
• Hajiagha MN, Kafil HS. Efflux pumps and microbial biofilm formation. Infect Genet Evol. 2023 Aug;112:105459. doi: 10.1016/j.meegid.2023.105459. Epub 2023 Jun 2. PMID 37271271
• Ren J, Wang M, Zhou W, Liu Z. Efflux pumps as potential targets for biofilm inhibition. Front Microbiol. 2024 Feb 23;15:1315238. doi: 10.3389/fmicb.2024.1315238. eCollection 2024. PMID 38596384
• Soto SM. Role of efflux pumps in the antibiotic resistance of bacteria embedded in a biofilm. Virulence. 2013 Apr 1;4(3):223-9. doi: 10.4161/viru.23724. Epub 2013 Feb 4. PMID 23380871

Идентификаторы

NCT: NCT07469436 · RBMEPGEEPAABFIMRCIOPA

Первоисточники (государственные реестры)

• ClinicalTrials.gov (NIH, США)

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