Investigating of four main carbapenem-resistance mechanisms in high-level carbapenem resistant Pseudomonas aeruginosa isolated from burn patients

Rostami, Soodabeh; Sheikh, Ahmad Farajzadeh; Shoja, Saeed; Farahani, Abbas; Tabatabaiefar, Mohammad Amin; Jolodar, Abbas; Sheikhi, Raheleh

doi:10.1016/j.jcma.2017.08.016

Cited by 58 publications

(51 citation statements)

References 23 publications

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“…39 Although the MexXY-OprM system is known as a substantial element in aminoglycoside resistance in only P. aeruginosa, 40 it is also over-expressed in multi-drug resistant P. aeruginosa 41 and similar reports to our study have shown its over-expression in carbapenemresistant P. aeruginosa strains. 7,42,43 MexAB-OprM over-expression was observed in 21.8% of the isolates, concurring with Xavier and co-workers. 44 In contrast, Dantas et al 7 and Chalhoub et al 32 reported a much higher frequency of MexAB-OprM over-expression in P. aeruginosa isolates.…”

Section: Discussionsupporting

confidence: 87%

“…Total DNA from P. aeruginosa isolates was extracted by boiling method according to Rostami et al 16 The tubes were stored at −20°C prior to being used in PCR amplification as a DNA template. 10 All carbapenem-resistant P. aeruginosa isolates were screened by PCR for the bla IMP1 , bla IMP2 , bla VIM1 , bla VIM2 , bla SPM and bla NDM genes using specific primers and conditions listed in Table 1.…”

Section: Dna Extraction and Pcrmentioning

confidence: 99%

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Molecular Epidemiology and Mechanisms of High-Level Resistance to Meropenem and Imipenem in Pseudomonas aeruginosa

Hassuna¹,

Darwish²,

Sayed³

et al. 2020

IDR

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Purpose: Pseudomonas aeruginosa possesses a large number of resistance mechanisms to different antimicrobials with carbapenems being the most powerful in treating resistant P. aeruginosa. Hence, it is imperative to explore different mechanisms of carbapenemsresistance in P. aeruginosa to achieve successful treatment through the design of new drugs acting on this interaction to combat against antimicrobial resistance. Strains and Methods: A total of 634 P. aeruginosa clinical isolates were collected from various patient sources and their MIC levels were measured. Molecular evaluation of carbapenem resistance was assessed by investigating the presence of bla IMP1 , bla IMP2 , bla VIM1 , bla VIM2 , bla SPM and bla NDM genes and the gene expression of the following multidrug efflux pump systems: MexAB-OprM, MexCD-OprJ, MexEF-OprN and MexXY-OprM and its correlation with MIC. Isolates were typed by Random Amplified Polymorphic DNA (RAPD)-typing. Results: Carbapenem resistance was detected in 32 (5%) isolates, which were all imipenem resistant (of which 29 were meropenem resistant). High-level resistance (≥64mg/mL) to imipenem was found in 27 (84.3%) isolates, and to meropenem in 28 (96.5%) isolates. The carbapenemase bla VIM-1 was found in 31 isolates, while bla NDM was detected in 4 isolates. None of the isolates possessed either bla-VIM-2 , bla IMP-1 , bla IMP-2 or bla SPM. The majority of the isolates displayed over-expression of MexCD-OprJ (75%) followed by MexXY-OprM efflux pump (62%), while MexAB-OprM and MexEF-OprN efflux pumps were overexpressed in 21.8% and 18.7% of the isolates, respectively, with no down-regulation of oprD in any of the isolates. A strong correlation was found between CDJ efflux pump expression and meropenem, imipenem resistance (r=0.532, 0.654, p<0.001, <0.001) respectively. Four major clusters were detected by RAPD-typing: group 1(10 isolates), group 3 (9 isolates), group 2 (8 isolates) while the fourth group (4) included 4 isolates (12.5% polymorphism). Conclusion: High-level carbapenem resistance reported in this study was allied to multiple mechanisms including carbapenemase production and efflux-pump over-expression. Threatening cross-infection is possible inside the hospital and stringent infection control measures are crucial.

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Section: Discussionsupporting

confidence: 87%

Section: Dna Extraction and Pcrmentioning

confidence: 99%

Molecular Epidemiology and Mechanisms of High-Level Resistance to Meropenem and Imipenem in Pseudomonas aeruginosa

Hassuna¹,

Darwish²,

Sayed³

et al. 2020

IDR

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“…On the other hand, more than 90% of the isolates with OprD downregulation showed an imipenem MIC value equal or more than 32 mg/L, and 86.4% of the isolates had a meropenem MIC ≥2 mg/L. However, while our data was in agreement with the results of Livermore [35], it wasn't in agreement with the results of Rostami [26] and Ellappan [36]. In addition, reduced expression (but not downregulation) or normal expression for oprD of some P. aeruginosa isolates were found, though these isolates showed high level resistance to imipenem in our study.…”

Section: Discussionsupporting

confidence: 53%

“…Genes encoding MBLs are transferred by mobile genetic elements [26]. MBL production is noted in massive outbreaks, including the international dissemination of resistant strains [27].…”

Section: Discussionmentioning

confidence: 99%

Role of efflux pump and OprD porin expression in carbapenem resistance of Pseudomonas aeruginosa clinical isolates

Müderris

Durmaz

Özdem

et al. 2018

J Infect Dev Ctries

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Introduction: In recent years, the prevalence of multidrug-resistant P. aeruginosa has remarkably increased. Thus, we wanted to investigate the carbapenem resistance mechanisms and clonal relationship among 80 carbapenem-resistant P. aeruginosa strains. Methodology: Carbapenemase production was detected using the Modified Hodge Test (MHT), EDTA combined disc method (ECD), and PCR. Expression levels of efflux and porin genes were mesured by real-time reverse transcription PCR. Clonal relationship of the isolates was investigated by pulsed-field gel electrophoresis (PFGE). Results: Carbapenemase production was detected in 7.5% of the isolates with MHT/ECD tests and in 11.3% of the isolates with PCR. Although the specificity of MHT/ECD was high, the sensitivitivity was low. oprD downregulation and mexB, mexY, and mexD overexpression were demonstrated in 55%, 16.3%, 2.5%, and 2.5% of the isolates, respectively. Multiple carbapenem resistance mechanisms were found in nearly a quarter of the isolates. PFGE typing of the 80 P. aeruginosa isolates yielded 61 different patterns. A total of 29 isolates (36.3%) were classified in 10 clusters, containing 2 to 7 strains. We could not find a strict relationship between PFGE profile and carbapenem resistance mechanisms. Conclusions: Although oprD downregulation and MexAB-OprM overexpression were the most common mechanisms, carbapenem resistance was associated with multiple mechanisms in the study. MHT/ECD tests should not be used alone for investigation of carbapenemase production in P. aeruginosa. Rapid tests with high sensitivity and specificity should be developed for the detection of carbapenemase production in P. aeruginosa.

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“…Studies have confirmed the involvement of genes previously proposed to be associated with resistance in P. aeruginosa. These include the fusA1 gene that encodes the EFG-1 protein and is associated with aminoglycoside resistance (27,29,30) and the ftsI gene that encodes a penicillin binding protein and is associated with carbapenem resistance (30)(31)(32)(33)(34). This approach has high potential for identifying previously unknown antibiotic resistanceassociated mutations and genes, but some key issues remain to be addressed.…”

Section: Introductionmentioning

confidence: 99%

A large-scale comparison shows that genetic changes causing antibiotic resistance in experimentally evolvedPseudomonas aeruginosapredict those in naturally evolved bacteria

Wardell¹,

Rehman²,

Martin³

et al. 2019

Preprint

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Pseudomonas aeruginosa is an opportunistic pathogen that causes a wide range of acute and chronic infections. An increasing number of isolates have acquired mutations that make them antibiotic resistant, making treatment more difficult. To identify resistance-associated mutations we experimentally evolved the antibiotic sensitive strain P. aeruginosa PAO1 to become resistant to three widely used anti-pseudomonal antibiotics, ciprofloxacin, meropenem and tobramycin. Mutants were able to tolerate up to 2048-fold higher concentrations of antibiotic than strain PAO1. Genome sequences were determined for thirteen mutants for each antibiotic. Each mutant had between 2 and 8 mutations. There were at least 8 genes mutated in more than one mutant per antibiotic, demonstrating the complexity of the genetic basis of resistance. Additionally, large deletions of up to 479kb arose in multiple meropenem resistant mutants. For all three antibiotics mutations arose in genes known to be associated with resistance, but also in genes not previously associated with resistance. To determine the clinical relevance of mutations uncovered in experimentallyevolved mutants we analysed the corresponding genes in 457 isolates of P. aeruginosa from patients with cystic fibrosis or bronchiectasis as well as 172 isolates from the general environment. Many of the genes identified through experimental evolution had changes predicted to be function-altering in clinical isolates but not in isolates from the general environment, showing that mutated genes in experimentally evolved bacteria can predict those that undergo mutation during infection. These findings expand understanding of the genetic basis of antibiotic resistance in P. aeruginosa as well as demonstrating the validity of experimental evolution in identifying clinically-relevant resistance-associated mutations.' ImportanceThe rise in antibiotic resistant bacteria represents an impending global health crisis. As such, understanding the genetic mechanisms underpinning this resistance can be a crucial piece of the puzzle to combatting it. The importance of this research is that by experimentally evolving P. aeruginosa to three clinically relevant antibiotics, we have generated a catalogue of genes that can contribute to resistance in vitro. We show that many (but not all) of these genes are clinically relevant, by identifying variants in clinical isolates of P. aeruginosa. This research furthers our understanding of the genetics leading to resistance in P. aeruginosa and provides tangible evidence that these genes can play a role clinically, potentially leading to new druggable targets or inform therapies.

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Investigating of four main carbapenem-resistance mechanisms in high-level carbapenem resistant Pseudomonas aeruginosa isolated from burn patients

Cited by 58 publications

References 23 publications

<p>Molecular Epidemiology and Mechanisms of High-Level Resistance to Meropenem and Imipenem in <em>Pseudomonas aeruginosa</em></p>

<p>Molecular Epidemiology and Mechanisms of High-Level Resistance to Meropenem and Imipenem in <em>Pseudomonas aeruginosa</em></p>

Role of efflux pump and OprD porin expression in carbapenem resistance of Pseudomonas aeruginosa clinical isolates

A large-scale comparison shows that genetic changes causing antibiotic resistance in experimentally evolvedPseudomonas aeruginosapredict those in naturally evolved bacteria

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