Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Jan-Mar;10(1):37-42.
doi: 10.4103/ijabmr.IJABMR_90_19. Epub 2020 Jan 3.

Detection of Overexpression of Efflux Pump Expression in Fluoroquinolone-Resistant Pseudomonas aeruginosa Isolates

Nouf Al Rashed  1 , Ronni Mol Joji  1 , Nermin Kamal Saeed  2 , Khalid Mubarak Bindayna  1
Affiliations

Detection of Overexpression of Efflux Pump Expression in Fluoroquinolone-Resistant Pseudomonas aeruginosa Isolates

Nouf Al Rashed et al. Int J Appl Basic Med Res. 2020 Jan-Mar.

Abstract

Context: Fluoroquinolones are the most effective antibiotics against Pseudomonas aeruginosa; many strains, however, have shown resistance due to mutations in DNA gyrase, topoisomerase IV, or in the efflux pumps. Little is known about P. aeruginosa efflux pump resistance mechanisms in the Kingdom of Bahrain.

Aim: The aim was to study efflux pump-mediated fluoroquinolone resistance among P. aeruginosa isolates using phenotypic (E-test and agar dilution) and genotypic (real-time-polymerase chain reaction [RT-PCR]) methods.

Materials and methods: Fifty ciprofloxacin-resistant P. aeruginosa isolates were included in this study. Genus and species of P. aeruginosa were confirmed by conventional PCR. The minimum inhibitory concentration (MIC) of ciprofloxacin with and without carbonyl cyanide 3-chlorophenylhydrazone (CCCP) was determined by E-test and agar dilution test. The overexpression of genes MexB, MexD, MexF, and MexY was measured by RT-PCR.

Results: All isolates were confirmed as P. aeruginosa. Among the fifty isolates, four showed reduction in ciprofloxacin MIC after addition of CCCP. These four isolates showed upregulation of expression of at least one of the four genes by RT-PCR. The mean gene expression of MexB, MexD, MexF, and MexY increased by 1.6, 4.65, 3.4, and 3.68-fold, respectively.

Conclusion: The results demonstrate the presence and type of efflux pump overexpression, mandating for large multicentric studies.

Keywords: Agar dilution; Pseudomonas aeruginosa; efflux pump; fluoroquinolone resistance; real-time-polymerase chain reaction.

PubMed Disclaimer

Conflict of interest statement

There are no conflicts of interest.

Figures

Figure 1
Figure 1
Polymerase chain reaction products on agarose gel electrophoresis of conventional polymerase chain reaction. Lanes (1–4) are two bands for isolate numbers 1–4, lane 5 is Pseudomonas aeruginosa ATCC27853 strain (positive control), lane 6 is water (negative control), and lane 7 is the ladder
Figure 2
Figure 2
Comparison of E-test strip result between plain Muller–Hinton Agar (a) and Muller–Hinton Agar with carbonyl cyanide 3-chlorophenylhydrazone (b) for isolate no. 16
Figure 3
Figure 3
Relative quantification results of the four isolates compared to ATCC27853 strain
See this image and copyright information in PMC

References

    1. Khan H, Ahmad A, Mehboob R. Nosocomial infections and their control strategies. Asian Pacific J Tropi Biomed. 2015;5:509.
    1. Hsueh PR, Tseng SP, Teng LJ, Ho SW. Pan-drug-resistant Pseudomonas aeruginosa causing nosocomial infection at a University hospital in Taiwan. Clin Microbiol Infect. 2005;11:670–3. - PubMed
    1. Pitt TL. Biology of Pseudomonas aeruginosa in relation to pulmonary infection in cystic fibrosis. J R Soc Med. 1986;79(Suppl 12):13–8. - PMC - PubMed
    1. Jami Al-Ahmadi G, Zahmatkesh Roodsari R. Fast and specific detection of Pseudomonas aeruginosa from other Pseudomonas species by PCR. Ann Burns Fire Disasters. 2016;29:264–7. - PMC - PubMed
    1. Algun U, Arisoy A, Gunduz T, Ozbakkaloglu B. The resistance of Pseudomonas aeruginosa strains to fluoroquinolone group of antibiotics. Indian J Med Microbiol. 2004;22:112–4. - PubMed