Comparation of Phenotypic and Genotypic Profile of Carbapenemase Producing Escherichia coli

Silvia Sutandhio, Budiono Budiono, Hardiono Hardiono, Kuntaman Kuntaman, Eddy Bagus Wasito, Maria Inge Lusida

= http://dx.doi.org/10.20473/fmi.v54i1.8045
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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

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References


Bajaj P, Singh NS, Virdi JS (2016). Escherichia coli -lactamase: What really matters. Front. Microbiol. http://dx.doi.org/10.3389/fmicb.2016.00417

BD (2011). BD phoenix automated microbiology system user’s manual. Maryland: Becton, Dickinson and Company

CLSI (2016). Performance standards for antimicrobial susceptibility testing; twenty-sixth informational supplement. Pennsylvania, Clinical and Laboratory Standards Institute

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 -lactamase by use of a boronic acid compound. J. Clin. Microbiol 46, 4083–4086

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

Galani I, Rekatsina PD, Hatzaki D, Plachouras D, Souli M, Giamarellou H (2008). Evaluation of different laboratory tests for the detection of Metallo--Lactamase production in Enterobacteriaceae. JAC 61, 548-553

Hattori T, Kawamura K, Arakawa Y (2013). Compari-son of test methods for detecting Metallo--Lactama-se-producing Gram-negative bacteria. Jpn. J. Infect. Dis 66, 512-518

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

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

Kim SY, Hong SG, Moland ES, Thomson KS (2007). Convenient test using a combination of chelating agents for detection of Metallo--Lactamases in the clinical laboratory. American Society for Microbio-logy. J. Clin. Microbiol 45, 2798-2801

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

Queenan AM, Bush K. 2007. Carbapenemases: the versatile ß-lactamases. Clin. Microbiol. Rev 20, 440-458

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

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

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

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

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

Thomson KS (2010). Extended-Spectrum--Lactamase, AmpC, and carbapenemase issues. J. Clin. Microbiol 48, 1019-1025

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

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


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