Relationship of OqxAB efflux pump to antibiotic resistance, mainly fluoroquinolones in Klebsiella pneumoniae, isolated from hospitalized patients

Document Type : Original Article

Authors

1 Department of Microbiology, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

2 Infectious Disease Research Center, Hamadan University of Medical Sciences, Hamadan, Iran

Abstract

Objective(s): This research was designed to study the prevalence of OqxAB efflux pump genes and also to investigate the relationship between efflux pump and resistance to antibiotics, especially to fluoroquinolones, evaluate the expression levels of OqxAB genes, and molecular typing of Klebsiella pneumoniae isolated from hospitalized patients in Hamadan hospitals, west of Iran.
Materials and Methods: In a cross-sectional study, 100 clinical strains of K. pneumoniae were isolated from hospitalized patients in three major teaching hospitals from January to June 2021. The antibiotic susceptibility of isolates was evaluated by the disk-diffusion agar method. The frequency of genes encoding oqxA and oqxB of efflux pump genes was investigated by PCR, and the expression of the oqxA efflux pump gene was investigated by the Real-time PCR method. The genetic relationship of K. pneumoniae isolates was analyzed by the Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR technique.
Results: According to our results, the multi-drug resistance phenotype (MDR) in 65% and high prevalence resistance to ciprofloxacin in 89% of K. pneumoniae isolates was detected. The higher prevalence of oqxA (95%) and oqxB (98%) was also detected. There was a significant relationship between ciprofloxacin resistance and the oqxB gene as well as between ceftriaxone and chloramphenicol resistance and the oqxA gene. The expression of the oqxA gene was higher in ciprofloxacin-resistant isolates.
Conclusion: The results of this study suggest a potential reservoir for the spread of OqxAB genes among hospital-acquired bacteria. Infection control strategies should be used prudently to reduce the spread of resistant strains of K. pneumoniae in hospitals.

Keywords


1. Onori R, Gaiarsa S, Comandatore F, Pongolini S, Brisse S, Colombo A, et al. Tracking nosocomial Klebsiella pneumoniae infections and outbreaks by whole-genome analysis: Small-scale Italian scenario within a single hospital. J Clin Microbiol 2015; 9:2861-2868.
2. Paczosa MK, Mecsas J. Klebsiella pneumoniae: Going on the offense with a strong defense. Microbiol Mol Biol Rev 2016; 80: 629–661.
3. Shadkam S, Goli HR, Mirzaei B, Gholami M, Ahanjan M. Correlation between antimicrobial resistance and biofilm formation capability among Klebsiella pneumoniae strains isolated from hospitalized patients in Iran. Ann Clin Microbiol Antimicrob 2021; 20:1-7.
4. Heidary M, Nasiri MJ, Dabiri H, Tarashi S. Prevalence of drug-resistant Klebsiella pneumoniae in Iran: A review article. Iran J Public Health 2018; 47:317.
5. Mirzaei B, Babaei R, Bazgir ZN, Goli HR, Keshavarzi S, Amiri E. Prevalence of Enterobacteriaceae spp. and its multidrug-resistant rates in clinical isolates: A two-center cross-sectional study. Mol Biol Rep 2021; 48:665-675.
6. Karimi K, Zarei O, Sedighi P, Taheri M, Doosti-Irani A, Shokoohizadeh L. Investigation of antibiotic resistance and biofilm formation in clinical isolates of Klebsiella pneumoniae. Int J Microbiol 2021; 14; 2021.
7. Farhadi M, Ahanjan M, Goli HR, Haghshenas MR, Gholami M. High frequency of multidrug-resistant (MDR) Klebsiella pneumoniae harboring several β-lactamase and integron genes collected from several hospitals in the north of Iran. Ann Clin Microbiol Antimicrob 2021; 20:1-9.
8. Galani I, Karaiskos I, Giamarellou H. Multidrug-resistant Klebsiella pneumoniae: mechanisms of resistance including updated data for novel β-lactam-β-lactamase inhibitor combinations. Expert Rev Anti Infect Ther 2021; 19:1457-1468.
9. Endimiani A, Luzzaro F, Perilli M, Lombardi G, Colì A, Tamborini A, et al. Bacteremia due to Klebsiella pneumoniae isolates producing the TEM-52 extended-spectrum β-lactamase: Treatment outcome of patients receiving imipenem or ciprofloxacin. Clin Infect Dis 2004; 38:243-251.
10. Andersson MI, MacGowan AP. Development of the quinolones. J Antimicrob Chemother 2003; 51: 1-11.
11. Martı´nez JL, Alonso A, Go´ mez-Go´ mez JM. Quinolone resistance by mutations in chromosomalgyrase genes just the tip of the iceberg? J Antimicrob Chemother 1998; 42: 683-688.
12. Tran JH, Jacoby GA, Hooper DC. Interaction of the plasmid-encoded quinolone resistance protein Qnr with Escherichia coli DNA gyrase. Antimicrob Agents Chemother 2005; 49: 118-25.
13. Nordmann P, Poirel L. Emergence of plasmidmediated resistance to quinolones in Enterobacteriaceae. J Antimicrob Chemother 2005; 56: 463-469.
14. Robicsek A, Jacoby GA, Hooper DC. The worldwide emergence of plasmid mediated quinolone resistance. Lancet Infect Dis 2006; 6: 629-640.
15. Li J, Zhang H, Ning J, Sajid A, Cheng G, Yuan Z, et al. The nature and epidemiology of OqxAB, a multidrug efflux pump. Antimicrob Resist Infect Control 2019; 8:44.
16. Hernando-Amado S, Blanco P, Alcalde-Rico M, Corona F, Reales-Calderon JA, Sanchez MB, et al. Multidrug efflux pumps as main players in intrinsic and acquired resistance to antimicrobials. Drug Resist Updat 2016; 28:13–27
17. Perez F, Rudin SD, Marshall SH, Coakley P, Chen L, Kreiswirth BN, et al. OqxAB, aquinolone and olaquindox efflux pump, is widely distributed among multidrug-resistant Klebsiella pneumoniae isolates of human origin. Antimicrob Agents Chemother 2013; 57: 4602-4603.
18. Bharatham N, Bhowmik P, Aoki M, Okada U, Sharma S, Yamashita E, et al. Structure and function relationship of OqxB efflux pump from Klebsiella pneumoniae. Nature Commun 2021; 12:1-2.
19. Van Belkum A TP, Dijkshoorn L, Haeggman S, Cookson B, Fry N, Fussing V, et al. Guidelines for the validation and application of typing methods for use in bacterial epidemiology. Clin Microbiol Infect 2007; 13:1-46.
20. Mahon C, Lehman D, Manuselis G. Textbook of Diagnostic Microbiology. 5th Edidtion. Missouri: Elsevier. 2014.
21. CLSI. Performance Standards for Antimicrobial Susceptibility Testing. 31st ed. CLSI supplement M100. Clinical and Laboratory Standards Institute; 2021.
22. Oliveira CF, Paim TG, Reiter KC, Rieger A, D’azevedo PA. Evaluation of four different DNA extraction methods in coagulase-negative Staphylococci clinical isolates. Rev Inst Med Trop Sao Paulo 2014; 56:29-33.    
23. Rodríguez-Martínez JM, Díaz de Alba P, Briales A, Machuca J, Lossa M, Fernández-Cuenca F, et al. Contribution of OqxAB efflux pumps to quinolone resistance in extended-spectrum-β-lactamase-producing Klebsiella pneumoniae. J Antimicrob Chemother 2013; 68:68-73.
24. Sedighi P, Zarei O, Karimi K, Taheri M, Karami P, Shokoohizadeh L. Molecular typing of Klebsiella pneumoniae clinical isolates by Enterobacterial repetitive intergenic consensus polymerase chain reaction. Int J Microbiol 2020;2020:1-5.
25. Moosavian M, Emam N. The first report of emerging mobilized colistin-resistance (mcr) genes and ERIC-PCR typing in Escherichia coli and Klebsiella pneumoniae clinical isolates in southwest Iran. Infect Drug Resist 2019; 12:1001–1010.
26. Madahiah BM, Noor US, Abdul S, Ali Abbas Q. Klebsiella pneumoniae urinary tract infections associated with long-term catheterization and spinal cord injuries. J Med Sci 2002; 2: 227-229.
27. Cepas V, López Y, Muñoz E, Rolo D, Ardanuy C, Martí S, et al. Relationship between biofilm formation and antimicrobial resistance in gram-negative bacteria. Microb Drug Resist 2019; 25:72-79.
28. Aditi Priyadarshini B, Mahalakshmi K, Naveen Kumar V. Mutant prevention concentration of ciprofloxacin against Klebsiella pneumoniae clinical isolates: An ideal prognosticator in treating multidrug-resistant strains. Int J Microbiol 2019; 2019:6850108.
29. Parsaie Mehr V, Shokoohizadeh L, Mirzaee M, Savari M. Molecular typing of Klebsiella pneumoniae isolates by enterobacterial repetitive intergenic consensus (ERIC)–PCR. Infect Epidemiol Microbiol 2017; 3:112-116.
30. Nirwati H, Sinanjung K, Fahrunissa F, Wijaya F, Napitupulu S, Hati VP, et al. Biofilm formation and antibiotic resistance of Klebsiella pneumoniae isolated from clinical samples in a tertiary care hospital, Klaten, Indonesia. BMC Proc 2019; 13: 1-8. 
31. Li J, Zhang H, Ning J, Sajid A, Cheng G, Yuan Z, Hao H. The nature and epidemiology of OqxAB, a multidrug efflux pump. Antimicrob Resist Infect Control 2019; 8:1-3.
32. Mohammadpour Bishak F, Ashrafi F, Moradi Bidhendi S, Mirzaie A, Noorbazargan H. The impact of Grammosciadium platycarpum Boiss. & Hausskn. extract on oqxA efflux pump gene expression in antibiotic resistant clinical isolates of Klebsiella pneumoniae using real time PCR. J Med Plants 2020; 19:291-304.
33. Taherpour A, Hashemi A. Detection of OqxAB efflux pumps, OmpK35 and OmpK36 porins in extended-spectrum-β-lactamase-producing Klebsiella pneumoniae isolates from Iran. Hippokratia 2013; 17:355.
34. Jamshidi MR, Zandi H, Eftekhar F. Correlation of quinolone-resistance, qnr genes and integron carriage in multidrug-resistant community isolates of Klebsiella spp. Iran J Basic Med Sci 2019; 22:1387-1391.
35. Xu Q, Jiang J, Zhu Z, Xu T, Sheng ZK, Ye M, et al. Efflux pumps AcrAB and OqxAB contribute to nitrofurantoin resistance in an uropathogenic Klebsiella pneumoniae isolate. Int J Antimicrob Agents 2019; 54:223-227.
36. Wong MH, Yan M, Chan EW, Biao K, Chen S. Emergence of clinical Salmonella enterica serovar typhimurium isolates with concurrent resistance to ciprofloxacin, ceftriaxone, and azithromycin. Antimicrob Agents Chemother 2014; 58:3752-3756.