Evaluation of the presence of AmpC (FOX) beta-lactamase gene in clinical strains of Escherichia coli isolated from hospitalized patients in Tabriz

Background and purpose: Beta-lactamase enzymes inactivate beta-lactam antibiotics by hydrolyzing the central nucleus. AmpC-type beta-lactamases hydrolyze cephalosporins, penicillins, and cephamycins. Therefore, the aim of this study was to determine antibiotic resistance and to investigate the presence of AmpC beta-lactamase gene in clinical strains of Escherichia coli isolated from isolated from hospitalized patients in Tabriz. Materials and Methods: In this cross-sectional descriptive study, 289 E. coli specimens were collected from clinical specimens. Disk diffusion method and combined disk method were used to determine the phenotype of extended spectrum β-Lactamase producing (ESBLs) strains. Then PCR was used to evaluate the presence of AmpC (FOX) beta-lactamase gene in the strains confirmed in phenotypic tests. Antibiotic resistance was also determined using disk diffusion by the Kibry-Bauer method. Results: A total of 121 isolates were identified as generators of beta-lactamase genes. 72 (59.5 %) isolates producing ESBL and 49 (40.5 %) isolates were identified as AmpC generators. In the PCR test, 31 isolates contained the FOX gene. The highest resistance was related to the antibiotics amoxicillin (76.12%), ceftazidime (70.24%) and nalidixic acid (65.05%). Conclusion: The results indicate an increase in the prevalence of beta-lactamase genes and increased resistance to beta-lactam antibiotics, which can be the result of improper use of antibiotics and not using antibiotic susceptibility tests before starting treatment. Also, using phenotypic and molecular diagnostic methods such as PCR together can be very useful.

Keywords:

Extended spectrum β-Lactamase producing, AmpC Escherichia coli, FOX gene,

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1. Eslami M, Najar Peerayeh S. Phenotypic and molecular detection of TEM, PER, and VEB beta-lactamases in clinical strains of Escherichia coli. Journal of Arak University of Medical Sciences. 2012;15(1):1-9.
2. Jafari Sales A, Mobaiyen H. Frequency and resistance patterns in clinical isolates of Escherichia coli Extended Spectrum Beta Lactamase producing treatment Centers in Marand city, Iran. New Cellular and Molecular Biotechnology Journal. 2017;7(26):19-26.
3. Jafari Sales A, Mobaiyen H, Farshbafi Nezhad Zoghi J, Nezamdoost Shadbad N, Purabdollah Kaleybar V. Antimicrobial resistance pattern of extended-spectrum β-Lactamases (ESBLs) producing Escherichia coli isolated from clinical samples in Tabriz city, Iran. Adv Environ Biol. 2014;8(16):179-82.
4. Jafari-Sales A, Rasi-Bonab F. Detection of the antibiotic resistance pattern in Escherichia coli isolated from urinary tract infections in Tabriz City. J Mol Microbiol. 2017;1(1):1-3.
5. Yazdi m, Nazemi A, MIR IM, Khataminejad M, Sharifi S, BABAI KM. Prevalence of SHV/CTX-M/TEM (ESBL) Beta-lactamase Resistance Genes in Escherichia Coli Isolated from Urinary Tract Infections in Tehran, Iran. 2010.
6. Agrawal P, Ghosh A, Kumar S, Basu B, Kapila K. Prevalence of extended-spectrum β-lactamases among Escherichia coli and Klebsiella pneumoniae isolates in a tertiary care hospital. Indian Journal of Pathology and Microbiology. 2008;51(1):139.
7. Jafari-Sales A. Study of Antibiotic Resistance and Prevalence of bla-TEM gene in Klebsiella pneumoniae Strains isolated from Children with UTI in Tabriz Hospitals. Focus On Medical Sciences Journal. 2018;4(1).
8. Jafari-Sales A, Bagherizadeh Y, Khalifehpour M, Abdoli-senejan M, Helali-Pargali R. Antibiotic resistance pattern and bla-TEM gene expression in Acinetobacter baumannii isolated from clinical specimens of Tabriz hospitals. Zanko Journal of Medical Sciences. 2019;20(65):20-9.
9. Jafari-Sales A, Khaneshpour H. Molecular Study of BlaIMP and BlaVIM Genes in Pseudomonas Aeruginosa Strains, Producer of Metallo Beta Lactamases Isolated from Clinical Samples in Hospitals and Medical Centers of Tabriz. Paramedical Sciences and Military Health. 2020;14(4):18-25.
10. Jafari-Sales A, Shadi-Dizaji A. Molecular analysis of CTX-M genes among ESBL producing in Pseudomonas aeru-ginosa isolated from clinical samples by Multiplex-PCR. Hozan J Environment Sci. 2018;2(5):17-29.
11. Peter-Getzlaff S, Polsfuss S, Poledica M, Hombach M, Giger J, Böttger E, et al. Detection of AmpC beta-lactamase in Escherichia coli: comparison of three phenotypic confirmation assays and genetic analysis. Journal of clinical microbiology. 2011;49(8):2924-32.
12. Rahimi M, Tajbakhsh M, Razaghi M, Tajeddin E, Alebouyeh M, Bazl M, et al. Frequency of β-lactamase producing isolates of Escherichia coli and their diversity in enzyme activities among the resistance isolates from patients with diarrhea and nosocomial infections in Tehran, Iran. Koomesh. 2014;15(2).
13. Shebani A, Aghaee S, Nasr R. Prevalence of gene TEM-1 in E. coli strains isolated from clinical samples in Damghan. J Islamic Azad Uni Microbial Biotech. 2010;3:15-22.
14. Mohamudha PR, Harish B, Parija S. Molecular description of plasmid-mediated AmpC β-lactamases among nosocomial isolates of Escherichia coli & Klebsiella pneumoniae from six different hospitals in India. The Indian journal of medical research. 2012;135(1):114.
15. Fleming P, Goldner M, Glass D. Observations on the Nature, Distribution, and Significance of Cephalosporinaae. Lancet. 1963:1399-401.
16. Philippon A, Arlet G, Jacoby GA. Plasmid-determined AmpC-type β-lactamases. Antimicrobial agents and chemotherapy. 2002;46(1):1-11.
17. Hanson ND, Sanders CC. Regulation of inducible AmpC beta-lactamase expression among Enterobacteriaceae. Current pharmaceutical design. 1999;5(11):881-94.
18. Lin C-F, Hsu S-K, Chen C-H, Huang J-R, Lo H-H. Genotypic detection and molecular epidemiology of extended-spectrum β-lactamase-producing Escherichia coli and Klebsiella pneumoniae in a regional hospital in central Taiwan. Journal of Medical Microbiology. 2010;59(6):665-71.
19. Clinical, Laboratory Standards Institute M. Performance standards for antimicrobial susceptibility testing. Twenty-third informational supplement. 2013.
20. Jafari M, Fallah F, Borhan RS, Navidinia M, KARIMI A, RAFII TS, et al. The first report of CMY, aac (6')-Ib and 16S rRNA methylase genes among Pseudomonas aeruginosa isolates from Iran. 2013.
21. ROSTAM ZAD A, PADERVAND AY. EVALUATION OF BLA-CTX-M-15 AND BLA-AMPC(FOX) BETA LACTAMASE GENES IN KLEBSIELLA PNEUMONIAE ISOLATES ISOLATED FROM PATIENTS IN ISFAHAN CITY HOSPITALS. JUNDISHAPUR SCIENTIFIC MEDICAL JOURNAL. 2016;15(6).
22. mansouri s, chitsaz m, HAJI HR, MIRZAEI M, GHEYNI M. Determination of resistance pattern of plasmid-mediated ampc β–lactamases producing isolate of Escherichia coli. 2009.
23. Pérez-Pérez FJ, Hanson ND. Detection of plasmid-mediated AmpC β-lactamase genes in clinical isolates by using multiplex PCR. Journal of clinical microbiology. 2002;40(6):2153-62.
24. den Drijver E, Verweij JJ, Verhulst C, Oome S, Soer J, Willemsen I, et al. Decline in AmpC β-lactamase-producing Escherichia coli in a Dutch teaching hospital (2013-2016). PLoS One. 2018;13(10):e0204864.
25. Kiiru J, Kariuki S, Goddeeris BM, Butaye P. Analysis of β-lactamase phenotypes and carriage of selected β-lactamase genes among Escherichia coli strains obtained from Kenyan patients during an 18-year period. BMC microbiology. 2012;12(1):155.
26. SOLTAN DM, Sabbaghi A, MOLLA AH, RASTEGAR LA, ESHRAGHIAN M, FALLAH MJ, et al. PREVALENCE OF AMPC AND SHV α-LACTAMASES IN CLINICAL ISOLATES OF ESCHERICHIA COLI FROM TEHRAN HOSPITALS. 2013.
27. Mansouri S, Neyestanaki DK, Shokoohi M, Halimi S, Beigverdi R, Rezagholezadeh F, et al. Characterization of AmpC, CTX-M and MBLs types of β-lactamases in clinical isolates of Klebsiella pneumoniae and Escherichia coli producing extended spectrum β-lactamases in Kerman, Iran. Jundishapur journal of microbiology. 2014;7(2).
28. Koshesh M, Mansouri S, Hashemizadeh Z, Kalantar-Neyestanaki D. Identification of extended-spectrum β-lactamase genes and ampc-β-lactamase in clinical isolates of escherichia coli recovered from patients with urinary tract infections in Kerman, Iran. Archives of Pediatric Infectious Diseases. 2017;5(2).
29. Deshpande LM, Jones RN, Fritsche TR, Sader HS. Occurrence of plasmidic AmpC type beta-lactamase-mediated resistance in Escherichia coli: report from the SENTRY Antimicrobial Surveillance Program (North America, 2004). Int J Antimicrob Agents. 2006;28(6):578-81.
30. Hussain M, Hasan F, Shah AA, Hameed A, Jung M, Rayamajhi N, et al. Prevalence of class A and AmpC b-lactamases in clinical Escherichia coli isolates from Pakistan Institute of Medical Science, Islamabad, Pakistan. Jpn J Infect Dis. 2011;64(3):249252.
31. Maleki A, Khosravi A, Ghafourian S, Pakzad I, Hosseini S, Ramazanzadeh R, et al. High prevalence of AmpC β-Lactamases in clinical isolates of Escherichia coli in Ilam, Iran. Osong public health and research perspectives. 2015;6(3):201-4.
32. Yazdi MKS, Dallal MMS, Mirzael H, Sabbaghi A, Mehrabadi JF, Lari AR, et al. Molecular detection of TEM broad spectrum β-lactamase in clinical isolates of Escherichia coli. African Journal of Biotechnology. 2011;10(46):9454-8.
33. Hasani A, Mohammadzadeh A, Kafil HS, Rezaee MA, Hasani A, Aghazadeh M. Characterization of TEM-, SHV-, CTX-and AmpC-type β-lactamases from cephalosporin resistant Escherichia coli isolates from Northwest of Iran. Journal of Pure and Applied Microbiology. 2015;9(4):3401-6.