Rapid Detection and Differentiation of KPC and MBL Carbapenemases among Enterobacterales Isolates by a Modified Combined-Disk Test

WEI, MING; WANG, PENG; WANG, SHUAI; YANG, CHUNXIA; GU, LI

doi:10.33073/pjm-2021-036

Abstract

This study was conducted to develop a cheap, rapid, and accurate modified combined-disk test (mCDT) approach to detect and differentiate KPC and MBL carbapenemases among clinical carbapenem-resistant Enterobacterales (CRE) isolates and simultaneously distinguish them from carbapenem-susceptible Enterobacterales (CSE) isolates. A total of 163 CRE and 90 third-generation cephalosporin-resistant Enterobacterales isolates were tested using imipenem and meropenem disks and different concentrations of carbapenemase inhibitors. The optimal sensitivity and specificity for detecting KPC carbapenemase were 97.2% and 100%, respectively. The sensitivity and specificity for detecting MBL carbapenemase were 100% and 100% with imipenem or meropenem and carbapenemase inhibitors within six hours. The inhibitory zone diameter of 18 mm for imipenem or meropenem disks without inhibitor could distinguish CRE from CSE isolates. Therefore, this mCDT approach may be a useful tool in clinical laboratories to detect CRE isolates and differentiate KPC and MBL producers, which is beneficial for patient management and hospital infection prevention and control.

References

Bassetti M, Giacobbe DR, Giamarellou H, Viscoli C, Daikos GL, Dimopoulos G, De Rosa FG, Giamarellos-Bourboulis EJ, Rossolini GM, Righi E, et al.; Critically Ill Patients Study Group of the European Society of Clinical Microbiology and Infectious Disease (ESCMID); Hellenic Society of Chemotherapy (HSC); Società Italiana di Terapia Antinfettiva (SITA). Management of KPC-producing Klebsiella pneumoniae infections. Clin Microbiol Infect. 2018 Feb;24(2):133–144. https://doi.org/10.1016/j.cmi.2017.08.030
Search in Google Scholar Back to article
Bogaerts P, Berger AS, Evrard S, Huang TD. Comparison of two multiplex immunochromatographic assays for the rapid detection of major carbapenemases in Enterobacterales. J Antimicrob Chemother. 2020 Jun 01;75(6):1491–1494. https://doi.org/10.1093/jac/dkaa043
Search in Google Scholar Back to article
Burckhardt I, Zimmermann S. Using matrix-assisted laser desorption ionization-time of flight mass spectrometry to detect carbapenem resistance within 1 to 2.5 hours. J Clin Microbiol. 2011 Sep; 49(9):3321–3324. https://doi.org/10.1128/JCM.00287-11
Search in Google Scholar Back to article
Carvalhaes CG, Picão RC, Nicoletti AG, Xavier DE, Gales AC. Cloverleaf test (modified Hodge test) for detecting carbapenemase production in Klebsiella pneumoniae: be aware of false positive results. J Antimicrob Chemother. 2010 Feb 01;65(2):249–251. https://doi.org/10.1093/jac/dkp431
Search in Google Scholar Back to article
CDC. Antibiotic resistance threats in the United States, 2019. Atlanta (USA): U.S. Department of Health and Human Services, Centers for Disease Control and Prevention; 2019. https://doi.org/10.15620/cdc:82532
Search in Google Scholar Back to article
CLSI. M100-S29 Performance standards for antimicrobial susceptibility testing. Wayne (USA): Clinical and Laboratory Standards Institute; 2019.
Search in Google Scholar Back to article
Cointe A, Walewski V, Hobson CA, Doit C, Bidet P, Dortet L, Bonacorsi S, Birgy A. Rapid carbapenemase detection with Xpert Carba-R V2 directly on positive blood vials. Infect Drug Resist. 2019 Oct;12:3311–3316. https://doi.org/10.2147/IDR.S204436
Search in Google Scholar Back to article
Doi Y, Potoski BA, Adams-Haduch JM, Sidjabat HE, Pasculle AW, Paterson DL. Simple disk-based method for detection of Klebsiella pneumoniae carbapenemase-type β-lactamase by use of a boronic acid compound. J Clin Microbiol. 2008 Dec;46(12):4083–4086. https://doi.org/10.1128/JCM.01408-08
Search in Google Scholar Back to article
Findlay J, Hopkins KL, Meunier D, Woodford N. Evaluation of three commercial assays for rapid detection of genes encoding clinically relevant carbapenemases in cultured bacteria. J Antimicrob Chemother. 2015 May;70(5):1338–1342. https://doi.org/10.1093/jac/dku571
Search in Google Scholar Back to article
Gutiérrez-Gutiérrez B, Salamanca E, de Cueto M, Hsueh PR, Viale P, Paño-Pardo JR, Venditti M, Tumbarello M, Daikos G, Cantón R, et al.; REIPI/ESGBIS/INCREMENT Investigators. Effect of appropriate combination therapy on mortality of patients with bloodstream infections due to carbapenemase-producing Enterobacteriaceae (INCREMENT): a retrospective cohort study. Lancet Infect Dis. 2017 Jul;17(7):726–734. https://doi.org/10.1016/S1473-3099(17)30228-1
Search in Google Scholar Back to article
Hecker SJ, Reddy KR, Totrov M, Hirst GC, Lomovskaya O, Griffith DC, King P, Tsivkovski R, Sun D, Sabet M, et al. Discovery of a cyclic boronic acid β-lactamase inhibitor (RPX7009) with utility vs class a serine carbapenemases. J Med Chem. 2015 May 14;58(9): 3682–3692. https://doi.org/10.1021/acs.jmedchem.5b00127
Search in Google Scholar Back to article
Jing X, Zhou H, Min X, Zhang X, Yang Q, Du S, Li Y, Yu F, Jia M, Zhan Y, et al. The simplified carbapenem inactivation method (sCIM) for simple and accurate detection of carbapenemase-producing gram-negative bacilli. Front Microbiol. 2018 Oct 30;9:2391. https://doi.org/10.3389/fmicb.2018.02391
Search in Google Scholar Back to article
Kieffer N, Poirel L, Nordmann P. Rapid immunochromatography-based detection of carbapenemase producers. Infection. 2019 Aug; 47(4):673–675. https://doi.org/10.1007/s15010-019-01326-1
Search in Google Scholar Back to article
King M, Heil E, Kuriakose S, Bias T, Huang V, El-Beyrouty C, McCoy D, Hiles J, Richards L, Gardner J, et al. Multicenter study of outcomes with ceftazidime-avibactam in patients with carbapenemresistant Enterobacteriaceae infections. Antimicrob Agents Chemother. 2017 Jul;61(7):e00449–e17. https://doi.org/10.1128/AAC.00449-17
Search in Google Scholar Back to article
Laolerd W, Akeda Y, Preeyanon L, Ratthawongjirakul P, Santanirand P. Carbapenemase-producing carbapenem-resistant Enterobacteriaceae from Bangkok, Thailand, and their detection by the Carba NP and modified carbapenem inactivation method tests. Microb Drug Resist. 2018 Sep;24(7):1006–1011. https://doi.org/10.1089/mdr.2018.0080
Search in Google Scholar Back to article
Lee AJ, Suh HS. Comparative evaluation of the modified carbapenem inactivation method for phenotypic detection of Guiana extended-spectrum β-lactamase-type carbapenemases in Enterobacterales. Lab Med. 2021 Apr 30:lmab026. https://doi.org/10.1093/labmed/lmab026
Search in Google Scholar Back to article
Li J, Li C, Cai X, Shi J, Feng L, Tang K, Tong Y, Li Y. Performance of modified carbapenem inactivation method and inhibitor-based combined disk test in the detection and distinguishing of carbapenemase producing Enterobacteriaceae. Ann Transl Med. 2019 Oct; 7(20):566. https://doi.org/10.21037/atm.2019.09.43
Search in Google Scholar Back to article
Livermore DM, Meunier D, Hopkins KL, Doumith M, Hill R, Pike R, Staves P, Woodford N. Activity of ceftazidime/avibactam against problem Enterobacteriaceae and Pseudomonas aeruginosa in the UK, 2015–16. J Antimicrob Chemother. 2018 Mar 01;73(3):648–657. https://doi.org/10.1093/jac/dkx438
Search in Google Scholar Back to article
Logan LK, Weinstein RA. The epidemiology of carbapenemresistant Enterobacteriaceae: the impact and evolution of a global menace. J Infect Dis. 2017 Feb 15;215(suppl_1):S28–S36. https://doi.org/10.1093/infdis/jiw282
Search in Google Scholar Back to article
Nordmann P, Dortet L, Poirel L. Carbapenem resistance in Enterobacteriaceae: here is the storm! Trends Mol Med. 2012 May;18(5): 263–272. https://doi.org/10.1016/j.molmed.2012.03.003
Search in Google Scholar Back to article
Papagiannitsis CC, Študentová V, Izdebski R, Oikonomou O, Pfeifer Y, Petinaki E, Hrabák J. Matrix-assisted laser desorption ionization-time of flight mass spectrometry meropenem hydrolysis assay with NH4HCO3, a reliable tool for direct detection of carbapenemase activity. J Clin Microbiol. 2015 May;53(5):1731–1735. https://doi.org/10.1128/JCM.03094-14
Search in Google Scholar Back to article
Petropoulou D, Tzanetou K, Syriopoulou VP, Daikos GL, Ganteris G, Malamou-Lada E. Evaluation of imipenem/imipenem +EDTA disk method for detection of metallo-β-lactamase-producing Klebsiella pneumoniae isolated from blood cultures. Microb Drug Resist. 2006 Mar;12(1):39–43. https://doi.org/10.1089/mdr.2006.12.39
Search in Google Scholar Back to article
Pfaller MA, Huband MD, Mendes RE, Flamm RK, Castanheira M. In vitro activity of meropenem/vaborbactam and characterisation of carbapenem resistance mechanisms among carbapenem-resistant Enterobacteriaceae from the 2015 meropenem/vaborbactam surveillance programme. Int J Antimicrob Agents. 2018 Aug;52(2):144–150. https://doi.org/10.1016/j.ijantimicag.2018.02.021
Search in Google Scholar Back to article
Pierce VM, Simner PJ, Lonsway DR, Roe-Carpenter DE, Johnson JK, Brasso WB, Bobenchik AM, Lockett ZC, Charnot-Katsikas A, Ferraro MJ, et al. Modified carbapenem inactivation method for phenotypic detection of carbapenemase production among Enterobacteriaceae. J Clin Microbiol. 2017 Aug;55(8): 2321–2333. https://doi.org/10.1128/JCM.00193-17
Search in Google Scholar Back to article
Poirel L, Walsh TR, Cuvillier V, Nordmann P. Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis. 2011 May;70(1):119–123. https://doi.org/10.1016/j.diagmicrobio.2010.12.002
Search in Google Scholar Back to article
Pournaras S, Zarkotou O, Poulou A, Kristo I, Vrioni G, Themeli-Digalaki K, Tsakris A. A combined disk test for direct differentiation of carbapenemase-producing Enterobacteriaceae in surveillance rectal swabs. J Clin Microbiol. 2013 Sep;51(9):2986–2990. https://doi.org/10.1128/JCM.00901-13
Search in Google Scholar Back to article
Queenan AM, Bush K. Carbapenemases: the versatile β-lactamases. Clin Microbiol Rev. 2007 Jul;20(3):440–458. https://doi.org/10.1128/CMR.00001-07
Search in Google Scholar Back to article
Tsai YM, Wang S, Chiu HC, Kao CY, Wen LL. Combination of modified carbapenem inactivation method (mCIM) and EDTA-CIM (eCIM) for phenotypic detection of carbapenemase-producing Enterobacteriaceae. BMC Microbiol. 2020 Dec;20(1):315. https://doi.org/10.1186/s12866-020-02010-3
Search in Google Scholar Back to article
Tsakris A, Poulou A, Pournaras S, Voulgari E, Vrioni G, Themeli-Digalaki K, Petropoulou D, Sofianou D. A simple phenotypic method for the differentiation of metallo-β-lactamases and class A KPC carbapenemases in Enterobacteriaceae clinical isolates. J Antimicrob Chemother. 2010 Aug 01;65(8):1664–1671. https://doi.org/10.1093/jac/dkq210
Search in Google Scholar Back to article
van Duin D, Doi Y. The global epidemiology of carbapenemase-producing Enterobacteriaceae. Virulence. 2017 May 19;8(4):460–469. https://doi.org/10.1080/21505594.2016.1222343
Search in Google Scholar Back to article
WHO. Prioritization of pathogens to guide discovery, research and development of new antibiotics for drug resistant bacterial infections, including tuberculosis [Internet]. Geneva (Switzerland): World Health Organization; 2017 [cited 2021 Apr 14 ]. Available from https://www.who.int/publications/i/item/WHO-EMP-IAU-2017.12
Search in Google Scholar Back to article
Yamada K, Sasaki M, Murakami H, Aoki K, Morita T, Ishii Y, Tateda K. Evaluation of the simplified carbapenem inactivation method as a phenotypic detection method for carbapenemase-producing Enterobacterales. J Microbiol Methods. 2021 Aug;187:106273. https://doi.org/10.1016/j.mimet.2021.106273
Search in Google Scholar Back to article
Zhang R, Liu L, Zhou H, Chan EW, Li J, Fang Y, Li Y, Liao K, Chen S. Nationwide surveillance of clinical carbapenem-resistant Enterobacteriaceae (CRE) strains in China. EBioMedicine. 2017 May; 19:98–106. https://doi.org/10.1016/j.ebiom.2017.04.032
Search in Google Scholar Back to article

Rapid Detection and Differentiation of KPC and MBL Carbapenemases among Enterobacterales Isolates by a Modified Combined-Disk Test

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