Download PDF
CC BY-NC-ND 4.0 · J Lab Physicians 2021; 13(02): 151-155
DOI: 10.1055/s-0041-1729472
Original Article

Evaluation of Two Phenotypic Methods for the Detection of Plasmid-Mediated AmpC β-Lactamases among Enterobacteriaceae Isolates

Ronni Mol P
1   Division of Microbiology (Medicine), Rajah Muthaiah Medical College, Annamalai University, Chidambaram, Tamil Nadu, India
2   Department of Microbiology, Immunology and Infectious Diseases, College of Medicine and Medical Sciences, Arabian Gulf University, Manama, Kingdom of Bahrain
,
Khalid Mubarak Bindayna
2   Department of Microbiology, Immunology and Infectious Diseases, College of Medicine and Medical Sciences, Arabian Gulf University, Manama, Kingdom of Bahrain
,
Ganesan Shanthi
1   Division of Microbiology (Medicine), Rajah Muthaiah Medical College, Annamalai University, Chidambaram, Tamil Nadu, India
› Author Affiliations
› Further Information
   Permissions and Reprints

Abstract

Objectives AmpC β-lactamases are cephalosporinases that confer resistance to cephalothin, cefazolin, cefoxitin, penicillin, and β-lactamase inhibitor-β-lactam combinations. Even though the AmpC resistance is reported, but the accurate occurrence of AmpC β-lactamases in Enterobacteriaceae members is still unknown. Techniques to identify AmpC producers are still evolving but not yet optimized for the clinical laboratory. Here we aimed to compare the test performance of two different phenotypic methods, that is inhibitor-based assay using boronic acid and disk approximation test for AmpC detection in Enterobacteriaceae isolates from a tertiary hospital microbiology laboratory.

Materials and Methods The study includes 137 nonrepeat Enterobacteriaceae strains. Bacterial isolates, that yielded a zone diameter of less than 18 mm for cefoxitin by disk diffusion method were considered potential AmpC producers and further confirmed by phenotype methods—inhibitor-based assay using boronic acid and disk approximation test. A multiplex polymerase chain reaction was used to detect the most common plasmid-mediated AmpC genes: ACC, FOX, MOX, DHA, CIT, and EBC.

Results Of the 137 clinical isolates, 58 (42.33%) were cefoxitin resistant, while 53.4 and 18.9% of the cefoxitin-resistant isolates were positive by inhibitor-based assay and disk approximation test. Multiplex PCR detected 42 (30.6%) isolates with AmpC genes. Of the 42 isolates, the inhibitor-based assay detected 25 (59.5%) isolates, while the disk approximation test detected nine (21.4%) isolates.

Conclusion Our findings suggest that inhibitor-based assay using boronic acid can be used for the detection of the isolates that harbor AmpC β-lactamases. This method is cost-effective, simple to perform, and easy to interpret. Thus AmpC detection as a routine in clinical laboratories can help in appropriate therapeutic intervention and improved infection control.

Keywords

Enterobacteriaceae - AmpC β-lactamases - inhibitor-based assay - disk approximation test - multiplex PCR


Publication History

Article published online:
15 June 2021

© 2021. The Indian Association of Laboratory Physicians. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial-License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/).

Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India

 

  • References

  • 1 Jacoby GA. AmpC beta-lactamases. Clin Microbiol Rev 2009; 22 (01) 161-182
    CrossrefPubMedGoogle Scholar
  • 2 Bush K, Jacoby GA. Updated functional classification of beta-lactamases. Antimicrob Agents Chemother 2010; 54 (03) 969-976
    CrossrefPubMedGoogle Scholar
  • 3 Tamma PD, Doi Y, Bonomo RA, Johnson JK, Simner PJ. Group ARL; Antibacterial Resistance Leadership Group. A Primer on AmpC β-lactamases: necessary knowledge for an increasingly multidrug-resistant world. Clin Infect Dis 2019; 69 (08) 1446-1455
    CrossrefPubMedGoogle Scholar
  • 4 Handa D, Pandey A, Asthana AK, Rawat A, Handa S, Thakuria B. Evaluation of phenotypic tests for the detection of AmpC beta-lactamase in clinical isolates of Escherichia coli. Indian J Pathol Microbiol 2013; 56 (02) 135-138
    CrossrefPubMedGoogle Scholar
  • 5 Mohd Khari FI, Karunakaran R, Rosli R, Tee Tay S. Genotypic and phenotypic detection of AmpC β-lactamases in Enterobacter spp. isolated from a teaching hospital in Malaysia. PLoS One 2016; 11 (03) 0150643
    CrossrefPubMedGoogle Scholar
  • 6 Gupta G, Tak V, Mathur P. Detection of AmpC lactamases in gram-negative bacteria. J Lab Physicians 2014; 6 (01) 1-6
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 Correa-Martínez CL, Idelevich EA, Sparbier K, Kostrzewa M, Becker K. Rapid detection of extended-spectrum β-lactamases (ESBL) and AmpC β-lactamases in Enterobacterales: development of a screening panel using the MALDI-TOF MS-based direct-on-target microdroplet growth assay. Front Microbiol 2019; 10 (13) 13
    CrossrefPubMedGoogle Scholar
  • 8 Yadav KK, Adhikari N, Khadka R, Pant AD, Shah B. Multidrug resistant Enterobacteriaceae and extended spectrum β-lactamase producing Escherichia coli: a cross-sectional study in National Kidney Center, Nepal. Antimicrob Resist Infect Control 2015; 4 (01) 42
    CrossrefPubMedGoogle Scholar
  • 9 Pal N, Hooja S, Sharma R, Maheshwari RK. Phenotypic detection and antibiogram of β-lactamase-producing Proteus species in a Tertiary Care Hospital, India. Ann Med Health Sci Res 2016; 6 (05) 267-273
    CrossrefPubMedGoogle Scholar
  • 10 Coudron PE. Inhibitor-based methods for detection of plasmid-mediated AmpC beta-lactamases in Klebsiella spp., Escherichia coli, and Proteus mirabilis. J Clin Microbiol 2005; 43 (08) 4163-4167
    CrossrefPubMedGoogle Scholar
  • 11 El-Hady SA, Adel LA. Occurrence and detection of AmpC β-lactamases among Enterobacteriaceae isolates from patients at Ain Shams University Hospital. Egypt J Med Hum Genet 2015; 16 (03) 239-244
    CrossrefGoogle Scholar
  • 12 Vanpelt E, Belkum AV, Hays JP. Principles and Technical Aspects of PCR Amplification. Netherlands:Springer 2008
  • 13 Marchese A, Arlet G, Schito GC, Lagrange PH, Philippon A. Characterization of FOX-3, an AmpC-type plasmid-mediated beta-lactamase from an Italian isolate of Klebsiella oxytoca. Antimicrob Agents Chemother 1998; 42 (02) 464-467
    CrossrefPubMedGoogle Scholar
  • 14 Thomson KS. Controversies about extended-spectrum and AmpC beta-lactamases. Emerg Infect Dis 2001; 7 (02) 333-336
    CrossrefPubMedGoogle Scholar
  • 15 Pérez-Pérez FJ, Hanson ND. Detection of plasmid-mediated AmpC beta-lactamase genes in clinical isolates by using multiplex PCR. J Clin Microbiol 2002; 40 (06) 2153-2162
    CrossrefPubMedGoogle Scholar
  • 16 Hariharan P, Bharani T, Franklyne JS, Biswas P, Solanki SS, Paul-Satyaseela M. Antibiotic susceptibility pattern of Enterobacteriaceae and non-fermenter gram-negative clinical isolates of microbial resource orchid. J Nat Sci Biol Med 2015; 6 (01) 198-201
    CrossrefPubMedGoogle Scholar
  • 17 Sarma JB, Bhattacharya PK, Kalita D, Rajbangshi M. Multidrug-resistant Enterobacteriaceae including metallo-β-lactamase producers are predominant pathogens of healthcare-associated infections in an Indian teaching hospital. Indian J Med Microbiol 2011; 29 (01) 22-27
    CrossrefPubMedGoogle Scholar
  • 18 Lazm AM, Al-Allak MH, Al-Asskar JA, Al-Dahmoshi HOM, Al-Khafaji NSK. Antibiotics resistance patterns among Enterobacteriaceae isolated from different clinical samples. Drug Invention Today 2019; 12 (05) 938-942
    Google Scholar
  • 19 Yilmaz NO, Agus N, Bozcal E, Oner O, Uzel A. Detection of plasmid-mediated AmpC β-lactamase in Escherichia coli and Klebsiella pneumoniae . Indian J Med Microbiol 2013; 31 (01) 53-59
    CrossrefPubMedGoogle Scholar
  • 20 Helmy MM, Wasfi R. Phenotypic and molecular characterization of plasmid mediated AmpC β-lactamases among Escherichia coli, Klebsiella spp., and Proteus mirabilis isolated from urinary tract infections in Egyptian hospitals. BioMed Res Int 2014; 2014: 171548
    CrossrefPubMedGoogle Scholar
  • 21 Tenover FC, Emery SL, Spiegel CA. et al. Identification of plasmid-mediated AmpC beta-lactamases in Escherichia coli, Klebsiella spp., and proteus species can potentially improve reporting of cephalosporin susceptibility testing results. J Clin Microbiol 2009; 47 (02) 294-299
    CrossrefPubMedGoogle Scholar