Spread of Efflux Pump Overexpressing-Mediated Fluoroquinolone Resistance and Multidrug Resistance in Pseudomonas aeruginosa by using an Efflux Pump Inhibitor
2
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
Background: Fluoroquinolone resistance in Pseudomonas aeruginosa may be due to efflux pump overexpression and/or target mutations. We designed this study to investigate the efflux pump mediated fluoroquinolone resistance and check the increasing effectiveness of fluoroquinolones in combination with an efflux pumps inhibitor among P. aeruginosa isolates from burn wounds infections. Materials and Methods: A total of 154 consecutive strains of P. aeruginosa were recovered from separate patients hospitalized in a burn hospital, Tehran, Iran. The isolates first were studied by disk diffusion antibiogram for 11 antibiotics and then minimum inhibitory concentration (MIC) experiments were performed to detect synergy between ciprofloxacin and the efflux pump inhibitor, carbonyl cyanide- m-chlorophenyl hydrazone (CCCP). Then to elucidate the inducing of multi drug resistance due to different efflux pumps activation in Fluoroquinolone resistant isolates, synergy experiments were also performed in random ciprofloxacin resistant isolates which have overexpressed efflux pumps phenotypically, using CCCP and selected antibiotics as markers for Beta-lactams and Aminoglycosides. The isolates were also tested by polymerase chain reaction (PCR) for the presence of the MexA, MexC and MexE, which encode the efflux pumps MexAB-OprM, MexCD-OprJ and MexEF-OprN. Results: Most of the isolates were resistant to 3 or more antibiotics tested. More than half of the ciprofloxacin resistant isolates exhibited synergy between ciprofloxacin and CCCP, indicating the efflux pump activity contributed to the ciprofloxacin resistance. Also increased susceptibility of random ciprofloxacin resistant isolates of P. aeruginosa to other selected antibiotics, in presence of CCCP, implied multidrug extrusion by different active efflux pump in fluoroquinolones resistant strains. All of Ciprofloxacin resistant isolates were positive for MexA, MexC and MexE genes simultaneously. Conclusion: In this burn hospital, where multidrug resistant P. aeruginosa isolates were prevalent, ciprofloxacin resistance and multidrug resistance due to the overexpression of fluoroquinolones mediated efflux pumps has also now emerged. Early recognition of this resistance mechanism should allow the use of alternative antibiotics and use an efflux pumps inhibitor in combination with antibiotic therapy.
Abstract
Background: Fluoroquinolone resistance in Pseudomonas aeruginosa may be due to efflux pump overexpression and/or target mutations. We designed this study to investigate the efflux pump mediated fluoroquinolone resistance and check the increasing effectiveness of fluoroquinolones in combination with an efflux pumps inhibitor among P. aeruginosa isolates from burn wounds infections. Materials and Methods: A total of 154 consecutive strains of P. aeruginosa were recovered from separate patients hospitalized in a burn hospital, Tehran, Iran. The isolates first were studied by disk diffusion antibiogram for 11 antibiotics and then minimum inhibitory concentration (MIC) experiments were performed to detect synergy between ciprofloxacin and the efflux pump inhibitor, carbonyl cyanide- m-chlorophenyl hydrazone (CCCP). Then to elucidate the inducing of multi drug resistance due to different efflux pumps activation in Fluoroquinolone resistant isolates, synergy experiments were also performed in random ciprofloxacin resistant isolates which have overexpressed efflux pumps phenotypically, using CCCP and selected antibiotics as markers for Beta-lactams and Aminoglycosides. The isolates were also tested by polymerase chain reaction (PCR) for the presence of the MexA, MexC and MexE, which encode the efflux pumps MexAB-OprM, MexCD-OprJ and MexEF-OprN. Results: Most of the isolates were resistant to 3 or more antibiotics tested. More than half of the ciprofloxacin resistant isolates exhibited synergy between ciprofloxacin and CCCP, indicating the efflux pump activity contributed to the ciprofloxacin resistance. Also increased susceptibility of random ciprofloxacin resistant isolates of P. aeruginosa to other selected antibiotics, in presence of CCCP, implied multidrug extrusion by different active efflux pump in fluoroquinolones resistant strains. All of Ciprofloxacin resistant isolates were positive for MexA, MexC and MexE genes simultaneously. Conclusion: In this burn hospital, where multidrug resistant P. aeruginosa isolates were prevalent, ciprofloxacin resistance and multidrug resistance due to the overexpression of fluoroquinolones mediated efflux pumps has also now emerged. Early recognition of this resistance mechanism should allow the use of alternative antibiotics and use an efflux pumps inhibitor in combination with antibiotic therapy.
참고문헌 (23)
- Askoura M, "Efflux pump inhibitors (EPIs) as new antimicrobial agents against Pseudomonas aeruginosa", Libyan J Med, 6, pp.5870~ (2011)
- Fagon JY, "Invasive and non-invasive strategies for management of suspected ventilator associated pneumonia", Ann Intern Med, 132, pp.621~630 (2000)
- Harris A, "Epidemiology and clinical outcomes of patients with multiresistant Pseudomonas aeruginosa", Clin Infect Dis, 28, pp.1128~1133 (1999)
- Hancock RE, "Resistance mechanisms in Pseudomonas aeruginosa and other non fermentative gram-negative bacteria", Clin Infect Dis, 27(Suppl 1), pp.S93~S99 (1998)
- Sefton AM, "Mechanisms of antimicrobial resistance: their clinical relevance in the new millennium", Drugs, 62, pp.557~566 (2002)
- Kriengkauykiat J, "Use of an efflux pump inhibitor to determine the prevalence of efflux pump-mediated fluoroquinolone resistance and multidrug resistance in Pseudomonas aeruginosa", Antimicrob Agents Chemother, 49, pp.565~570 (2005)
- Köhler T, "Differential selection of multidrug efflux systems by quinolones in Pseudomonas aeruginosa", Antimicrob Agents Chemother, 41, pp.2540~2543 (1997)
- Chuanchuen R, "The MexJK efflux pump of Pseudomonas aeruginosa requires OprM for antibiotic efflux but not for efflux of triclo", J Bacteriol, 184, pp.5036~5044 (2002)
- Li Y, "A new member of the tripartite multidrug efflux pumps, MexVW-OprM, in Pseudomonas aeruginosa", J Antimicrob Chemother, 52, pp.572~575 (2003)
- Poole K, "Efflux-mediated resistance to fluoroquinolones in gram negative bacteria", Antimicrob Agents Chemother, 44, pp.2233~2241 (2000)
- Gotoh N, "Characterization of the MexC-MexD-OprJ multidrug efflux system in DeltamexA-mexB-oprM mutants of Pseudomonas aeruginosa", Antimicrob Agents Chemother, 42, pp.1938~1943 (1998)
- Schweizer H.P, "Efflux as a mechanism of resistance to antimicrobials in Pseudomonas aeruginosa and related bacteria: unanswered question", Genet Mol Res, 2, pp.48~62 (2003)
- Ikonomidis A, "Effect of the proton motive force inhibitor carbonyl cyanide-m-chlorophenylhydrazone (CCCP) on Pseudomonas aeruginosa biofilm development", Lett Appl Microbiol, 47, pp.298~302 (2008)
- Pagès JM, "Inhibitors of efflux pumps in Gram-negative bacteria", Trends Mol Med, 11, pp.382~389 (2005)
- Barrow G, "Characters of gram-negative bacteria in Cowan & steel manual for identification of medical bacteria", pp.130~131 (2003)
- Vos D, "Direct detection and identification of Pseudomonas aeruginosa in clinical samples such as skin biopsy specimens and expectorations by multiplex PCR based on two outer membrane lipoprotein genes, oprI and oprL", J Clin Microbiol, 35, pp.1295~1299 (1997)
- Hancock RE, "Outer membranes of gram-negative bacteria. XIX. Isolation from Pseudomonas aeruginosa PAO1 and use in reconstitution and definition of the permeability barrier", J Bacteriol, 136, pp.381~390 (1978)
- Pournaras S, "Spread of efflux pump-overexpressing, non-metallo-beta-lactamase-producing, meropenem-resistant but ceftazidime-susceptible Pseudomonas aeruginosa in a region with blaVIM endemicity", J Antimicrob Chemother, 56, pp.761~764 (2005)
- Bonfiglio G, "Antibiotic resistance in Pseudomonas aeruginosa: an Italian survey", J Antimicrob Chemother, 41, pp.307~310 (1998)
- Clinical and Laboratory Standards Institute, "Performance standards for antimicrobial susceptibility testing; twenty-second informational supplement", CLSI, ~ (2010)
- Masuda N, "Substrate specificities of MexAB-OprM, Mex-CD-OprJ, and MexXY-OprM efflux pumps in Pseudomonas aeruginosa", Antimicrob Agents Chemother, 44, pp.3322~3327 (2000)
- Quale J, "Molecular epidemiology and mechanisms of carbapenem resistance in Acinetobacter baumannii endemic in New York City", Clin Infect Dis, 37, pp.214~220 (2003)
- Lomovskaya O, "Identification and characterization of inhibitors of multidrug resistance efflux pumps in Pseudomonas aeruginosa: novel agents for combination therapy", Antimicrob Agents Chemother, 45, pp.105~116 (2001)