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Abstract

Carbapenemase-producing Escherichia coli (E. coli) has caused trouble in therapeutic antibiotic selection. Carbapenemase screening procedure in laboratories is usually based on inacurate semi-automatic system. Confirmation and classification of carbapenemases according to Ambler can be done with combination of phenotypic methods, i.e., Modified Hodge Test (MHT), Sodium Mercaptoacetic Acid (SMA), and 3-Aminophenylboronic Acid (PBA). This study aimed to compare profiles of carbapenemase-producing E. coli which were confirmed and classified phenotypically with the genotypic profiles. E. coli isolates from urine specimens which were potential as carbapenemase-producers according to semi-automatic system BD Phoenix were phenotypically tested with MHT, SMA, and PBA. Isolates were grouped as carbapenemase-producers and non carbapenemase-producers. Phenotypic carbapenemase-producer isolates were classified based on Ambler criteria. All isolates were then tested with Polymerase Chain Reaction (PCR) for the presence of OXA-48, IMP1, IMP2, GES, VIM, NDM, KPC genes. Out of 30 isolates, 6 isolates (20.0%) were MHT positive, and 25 isolates (83.3%) were SMA positive, which indicated that most isolates produced were carbapenemase Ambler B. PCR confirmed 12 isolates (40.0%) had VIM gene which were classified as carbapenemase Ambler B. Phenotypic confirmatory test had 100% sensitivity and 22.2% specificity. Classification with phenotypic confirmatory test had 91.7% match with PCR. Phenotypic confirmatory test detected more carbapenemase than PCR. This low specificity may be caused by inappropriate use of diagnostic gold standard. PCR should not be used for routine carbapenemase confirmation because of vast diversity of carbapenemases. Phenotypic confirmatory test can classify carbapenemase according to Ambler classification.

Keywords

Escherichia coli carbapenemase phenotypic genotypic

Article Details

How to Cite
Sutandhio, S., Budiono, B., Hardiono, H., Kuntaman, K., Wasito, E. B., & Lusida, M. I. (2018). Comparation of Phenotypic and Genotypic Profile of Carbapenemase Producing Escherichia coli. Folia Medica Indonesiana, 54(1), 10–15. https://doi.org/10.20473/fmi.v54i1.8045

References

  1. Bajaj P, Singh NS, Virdi JS (2016). Escherichia coli ï¢-lactamase: What really matters. Front. Microbiol. http://dx.doi.org/10.3389/fmicb.2016.00417
  2. BD (2011). BD phoenix automated microbiology system user's manual. Maryland: Becton, Dickinson and Company
  3. CLSI (2016). Performance standards for antimicrobial susceptibility testing; twenty-sixth informational supplement. Pennsylvania, Clinical and Laboratory Standards Institute
  4. Doi Y, Potoski BA, Adams-Haduch JM, Sidjabat HE, Pasculle AW, Paterson DL (2008). Simple disk-based method for detection of Klebsiella pneumoniae Carbapenemase-type ï¢-lactamase by use of a boronic acid compound. J. Clin. Microbiol 46, 4083–4086
  5. EUCAST (2013). EUCAST guidelines for detection of resistance mechanisms and specific resistances of clinical and/or epidemiological importance version 1.0. Växjö, European Committee on Antimicrobial Susceptibility Testing
  6. Galani I, Rekatsina PD, Hatzaki D, Plachouras D, Souli M, Giamarellou H (2008). Evaluation of different laboratory tests for the detection of Metallo-ï¢-Lactamase production in Enterobacteriaceae. JAC 61, 548-553
  7. Hattori T, Kawamura K, Arakawa Y (2013). Compari-son of test methods for detecting Metallo-ï¢-Lactama-se-producing Gram-negative bacteria. Jpn. J. Infect. Dis 66, 512-518
  8. Hsu LY, Apisarnthanarak A, Khan E, Suwantarat N, Ghafur A, Tambyah PA (2017). Carbapenem-resistant Acinetobacter baumannii and Enterobacteriaceae in South and Southeast Asia. Clin. Microbiol. Rev 30, 1-22
  9. Karuniawati A, Saharman YR, Lestari DC. 2013. Detec-tion of carbapenemase encoding genes in Enterobac-teriaceae, Pseudomonas aeruginosa, and Acineto-bacter baumannii isolated from patients at Intensive Care Unit Cipto Mangunkusumo Hospital in 2011. Acta Medica Indonesiana – The Indonesian Journal of Internal Medicine 45, 101-106
  10. Kim SY, Hong SG, Moland ES, Thomson KS (2007). Convenient test using a combination of chelating agents for detection of Metallo-ï¢-Lactamases in the clinical laboratory. American Society for Microbio-logy. J. Clin. Microbiol 45, 2798-2801
  11. Pournaras S, Poulou A, Tsakris A. 2010. Inhibitor-based methods for the detection of KPC-carbapenemase producing Enterobacteriaceae in clinical practice by using boronic acid compounds. J. Antimicrob. Chemother 65, 1319-1321
  12. Queenan AM, Bush K. 2007. Carbapenemases: the versatile íŸ-lactamases. Clin. Microbiol. Rev 20, 440-458
  13. Ribeiro VB, Linhares AR, Zavascki AP, Barth AL (2014). Performance of quantification of Modified Hodge Test: an evaluation with Klebsiella pneumo-niae carbapenemase-producing Enterobacteriaceae isolates. BioMed Research International
  14. Shenoy KA, Jyothi EK, Ravikumar R (2014). Phenotypic identification & molecular detection of blaNDM-1 gene in multidrug resistant Gram-negative bacilli in a tertiary care centre. Indian J Med Res 139, 625-631
  15. Stuart JC, Leverstein-Van Hall MA (2010). Guideline for phenotypic screening and confirmation of carbapenemases in Enterobacteriaceae. International Journal of Antimicrobial Agents 36, 205–210
  16. Sutandhio S, Alimsardjono L, Lusida MI (2015). Distribusi dan pola kepekaan Enterobacteriaceae dari spesimen urin di RSUD Dr. Soetomo Surabaya periode Januari – Juni 2015. Jurnal Widya Medika Surabaya 3, 53-59
  17. Tängdén T (2012). Multidrug-resistant Escherichia coli and Klebsiella pneumoniae: treatment, selection and international spread. Acta Universitatis Upsaliensis. Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine 840 72
  18. Thomson KS (2010). Extended-Spectrum-ï¢-Lactamase, AmpC, and carbapenemase issues. J. Clin. Microbiol 48, 1019-1025
  19. Tsakris A, Themeli-Digalaki K, Poulou A, Vrioni G, Voulgari E, Koumaki V, Agodi A, Pournaras S, Sofianou D (2011). Comparative evaluation of combined-disk tests using different boronic acid compounds for detection of Klebsiella pneumoniae carbapenemase-producing Enterobacteriaceae clinical isolates. JCM 49, 2804-2809
  20. Woodford N, Eastaway AT, Ford M, Leanord A, Keane C, Quayle RM, Steer JA, Zhang J, Livermore DM (2010). Comparison of BD Phoenix, Vitek 2, and MicroScan automated systems for detection and inference of mechanisms responsible for carbapenem resistance in Enterobacteriaceae. J. Clin. Microbiol 48, 2999-3002