Evaluation of Phenotypic and Genotypic Carbapenemase Genes in Gram-Negative Bacteria Resistant to Carbapenem and Determining their Antibiotic Resistance

Document Type : Original Article (s)

Authors

1 PhD Student, Department of Medical Bacteriology, Division of Microbiology, Pasteur Institute of Iran, Tehran, Iran

2 Department of Microbiology, Saveh Research and Science Branch, Islamic Azad University, Saveh, Iran

3 Professor, Department of Microbiology, Division of Microbiology, Pasteur Institute of Iran, Tehran, Iran

Abstract

Background: Carbapenem antibiotics are often used as the last line of treatment for infections caused by resistant Gram-negative bacteria (GNB). The most important mechanism of resistance to carbapenems is production of carbapenemase. The prevalence of carbapenem-resistant Gram-negative bacteria is on the rise worldwide, posing a major public health threat. The aim of this study was to determine the antibiotic resistance patterns and to detect carbapenemase producing Gram-negative bacteria in clinical isolates.Methods: In this study, a total of 134 clinical isolates of Gram-negative bacteria were collected in Tehran city, Iran, during 2012-2013. Antimicrobial susceptibility testing was performed and resistance genes were characterized using polymerase chain reaction (PCR) amplification and sequencing. The modified Hodge test (MHT) and ethylenediaminetetraacetic acid (EDTA) double-disk diffusion test (DDST) were performed for the screening carbapenemases.Findings: The number and frequency of isolated bacteria were as Escherichia coli 57 (42.5%), Klebsiella pneumonia 26 (19.4%), Acinetobacter spp 21 (15.6%), Pseudomonas aeruginosa 17 (12.6%), Citrobacter spp 8 (5.9%), Proteus spp 3 (2.2%), and Enterobacter spp 2 (1.4%). These organisms showed the highest resistance to ertapenem, cefotaxime, aztreonam, ceftazidime, amikacin and kanamaycin (100%). Confirmatory tests showed that 44 isolates (32.8%) were DDST positive and 17 isolates (12.6%) were MHT positive. Most of MHT-positive (41.17%) and DDST-positive (39.13%) isolates were Acinetobacter spp. The PCR based screening revealed the presence of the blaIMP-1 gene in 4 isolates.Conclusion: Owing to the presentation of blaIMP-1 gene in Klebsiella, Citrobacter, Acinetobacter and Escherichia coli and feasibility of horizontal gene transfer among bacteria, changing in antibiotic prescription policies is required. Due to the importance of Metallo beta lactamase producting strains in hospital, early identification could play an effective role in prevention and treatment of these isolates.

Keywords

References

  1. Bell BG, Schellevis F, Stobberingh E, Goossens H, Pringle M. A systematic review and meta-analysis of the effects of antibiotic consumption on antibiotic resistance. BMC Infect Dis 2014; 14: 13.
  2. Bush K, Jacoby GA. Updated functional classification of beta-lactamases. Antimicrob Agents Chemother 2010; 54(3): 969-76.
  3. Nordmann P, Dortet L, Poirel L. Carbapenem resistance in Enterobacteriaceae: here is the storm! Trends Mol Med 2012; 18(5): 263-72.
  4. Blair JM, Webber MA, Baylay AJ, Ogbolu DO, Piddock LJ. Molecular mechanisms of antibiotic resistance. Nat Rev Microbiol 2015; 13(1): 42-51.
  5. Clinical and Laboratory Standards Institute (CLSI). Performance standards for antimicrobial disk susceptibility tests. Approved standard M2-A9. 9th ed. Wayne PA; CLSI: 2006.
  6. Saito R, Koyano S, Dorin M, Higurashi Y, Misawa Y, Nagano N, et al. Evaluation of a simple phenotypic method for the detection of carbapenemase-producing Enterobacteriaceae. J Microbiol Methods 2015; 108: 45-8.
  7. Mochon AB, Garner OB, Hindler JA, Krogstad P, Ward KW, Lewinski MA, et al. New Delhi metallo-beta-lactamase (NDM-1)-producing Klebsiella pneumoniae: case report and laboratory detection strategies. J Clin Microbiol 2011; 49(4): 1667-70.
  8. Salabi AE, Toleman MA, Weeks J, Bruderer T, Frei R, Walsh TR. First report of the metallo-beta-lactamase SPM-1 in Europe. Antimicrob Agents Chemother 2010; 54(1): 582.
  9. Shahcheraghi F, Abbasalipour M, Feizabadi M, Ebrahimipour G, Akbari N. Isolation and genetic characterization of metallo-beta-lactamase and carbapenamase producing strains of Acinetobacter baumannii from patients at Tehran hospitals. Iran J Microbiol 2011; 3(2): 68-74.
  10. Shahcheraghi F, Nobari S, Rahmati GF, Nasiri S, Owlia P, Nikbin VS, et al. First report of New Delhi metallo-beta-lactamase-1-producing Klebsiella pneumoniae in Iran. Microb Drug Resist 2013; 19(1): 30-6.
  11. Osano E, Arakawa Y, Wacharotayankun R, Ohta M, Horii T, Ito H, et al. Molecular characterization of an enterobacterial metallo beta-lactamase found in a clinical isolate of Serratia marcescens that shows imipenem resistance. Antimicrob Agents Chemother 1994; 38(1): 71-8.
  12. Yousefi S, Farajnia S, Nahaei MR, Akhi MT, Ghotaslou R, Soroush MH, et al. Detection of metallo-beta-lactamase-encoding genes among clinical isolates of Pseudomonas aeruginosa in northwest of Iran. Diagn Microbiol Infect Dis 2010; 68(3): 322-5.
  13. Feizabadi MM, Fathollahzadeh B, Taherikalani M, Rasoolinejad M, Sadeghifard N, Aligholi M, et al. Antimicrobial susceptibility patterns and distribution of blaOXA genes among Acinetobacter spp. Isolated from patients at Tehran hospitals. Jpn J Infect Dis 2008; 61(4): 274-8.
  14. Pena I, Picazo JJ, Rodriguez-Avial C, Rodriguez-Avial I. Carbapenemase-producing Enterobacteriaceae in a tertiary hospital in Madrid, Spain: high percentage of colistin resistance among VIM-1-producing Klebsiella pneumoniae ST11 isolates. Int J Antimicrob Agents 2014; 43(5): 460-4.
  15. Tangden T, Giske CG. Global dissemination of extensively drug-resistant carbapenemase-producing Enterobacteriaceae: clinical perspectives on detection, treatment and infection control. J Intern Med 2015; 277(5): 501-12.
  16. Tenover FC, Kalsi RK, Williams PP, Carey RB, Stocker S, Lonsway D, et al. Carbapenem resistance in Klebsiella pneumoniae not detected by automated susceptibility testing. Emerg Infect Dis 2006; 12(8): 1209-13.
  17. Azimi L, Nordmann P, Lari AR, Bonnin RA. First report of OXA-48-producing Klebsiella pneumoniae strains in Iran. GMS Hyg Infect Control 2014; 9(1): Doc07.
Journal of Isfahan Medical School
Volume 34, Issue 405 - Serial Number 405
September and October 2017
Pages 1290-1296

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History

  • Receive Date: 05 October 2015
  • Revise Date: 07 June 2024
  • Accept Date: 06 April 2022

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