Evaluation of Combined Use of Temocillin Disk and Mastdisks Inhibitor Combination Set Against Polymerase Chain Reaction for Detection of Carbapenem-Resistant Enterobacteriaceae

Authors

  • Omnia A. Shaker Clinical Pathology Department - Faculty of Medicine - Ain Shams University, Cairo
  • Howayda Ezz El Din Gomaa Clinical Pathology Department -National Research Centre - Egypt, Cairo http://orcid.org/0000-0002-3444-5898
  • Shereen A. ElMasry Clinical Pathology Department - Faculty of Medicine - Ain Shams University, Cairo
  • Rania M. Abdel Halim Clinical Pathology Department - Faculty of Medicine - Ain Shams University, Cairo
  • Amany H. Abdelrahman Clinical Pathology Department -National Research Centre - Egypt, Cairo
  • Jackline S. Kamal Clinical Pathology Department -National Research Centre - Egypt, Cairo

DOI:

https://doi.org/10.3889/oamjms.2018.090

Keywords:

Enterobacteriaceae, Carbapenemases, Mastdisks inhibitor, Temocillin

Abstract

AIMS: To evaluate the diagnostic performance of MDI and temocillin disk (30 μg) for detection of carbapenem-resistant Enterobacteriaceae in comparison to real-time PCR.

METHODS AND RESULTS: Fifty specimens submitted to the Microbiology Laboratory of Ain Shams University Hospitals and showed resistance to carbapenem drugs through routine culture and susceptibility testing, were assessed by both temocillin disk (30 μg) and MDI set to detect carbapenem-resistant Enterobacteriaceae. Results were compared to real-time PCR for detection of carbapenemase genes blaKPC, blaNDM, blaOXA–48-like, blaVIM, and blaIMP. Our work revealed that most of the CPE isolates were Klebsiella species (62%) followed by E. coli (24%), Serratia (10%) and Citrobacter (4%). Phenotypic detection of carbapenem-resistant classes revealed OXA - 48 in 96% of isolates, followed by MBLs (82%), and KPC (34%).  All isolates were negative for AmpC.  Detection of the genes by real-time PCR showed that the predominance was for the blaOXA-48 gene (96%) then blaVIM (94%) followed by blaNDM (54%), blaKPC (46%) and finally blaIMP (40%). Evaluation of the MDI set against PCR showed sensitivity (82.1%) and specificity (70%). The temocillin disk had 97.9% sensitivity and 50% specificity.  The evaluation of Temocillin disk and MDI in combination for detection of carbapenem-resistant Enterobacteriaceae showed 99.7% sensitivity and 35% specificity.

CONCLUSIONS: Adding Temocillin disk to Mastdisks ID inhibitor combination set provides a simple, easy, rapid and highly sensitive test that can be used for screening and classification of carbapenem-resistant Enterobacteriaceae. However, it still needs confirmation by molecular techniques.

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References

Singh-Moodley A, Perovic. Antimicrobial susceptibility testing in predicting the presence of carbapenemase genes in Enterobacteriaceae in South Africa. BMC Infect Dis. 2016; 16: 536. https://doi.org/10.1186/s12879-016-1858-7 PMid:27716102 PMCid:PMC5050574

Queenan AM, Bush K. Carbapenemases the versatile beta-lactamases. Clin Microbiol Rev. 2007; 20(3):440–458. https://doi.org/10.1128/CMR.00001-07 PMid:17630334 PMCid:PMC1932750

Falagas M, Lourida P, Poulikakos P, et al.: Antibiotic treatment of infections due to carbapenem-resistant Enterobacteriaceae: a systematic evaluation of the available evidence. Antimicrob Agents Chemother. 2014; 58(2):654-663. https://doi.org/10.1128/AAC.01222-13 PMid:24080646 PMCid:PMC3910850

Fernández-Cuenca F, Rodríguez-Martínez JM, Gómez-Sánchez MA, et al. Production of a plasmid-encoded OXA-72 β-lactamase associated with resistance to carbapenems in a clinical isolate Acinetobacter junii. Int J Antimicrob Agents. 2012; 39(1):93–94. https://doi.org/10.1016/j.ijantimicag.2011.07.017 PMid:21982835

Agila K. Pragasam, Sahni D. Rani, Anandan S, et al. A Pilot Study on Carbapenemase Detection: Do We See the Same Level of Agreement as with the CLSI Observations. Journal of Clinical and Diagnostic Research. 2016; 10(7): DC09-DC13.

Walther-Rasmussen J and Hoiby N. Class A carbapenemases. J Antimicrob Chemother. 2007; 60:470–482. https://doi.org/10.1093/jac/dkm226 PMid:17595289

Nordmann P, Naas T and Poirel L. Global spread of carbapenemase-producing Enterobacteriaceae. Emerg Infect Dis. 2011; 17:1791–1798. https://doi.org/10.3201/eid1710.110655 PMid:22000347 PMCid:PMC3310682

Girmenia C, Serrao A, and Canichella M. Epidemiology of Carbapenem Resistant Klebsiella pneumoniae Infections in Mediterranean Countries. Mediterr J Hematol Infect Dis. 2016; 8(1): e2016032. https://doi.org/10.4084/mjhid.2016.032 PMid:27441063 PMCid:PMC4943068

Poirel L, Potron A, and Nordmann P. OXA-48-like carbapenemases: the phantom menace. J Antimicrob Chemother, 2012; 67: 1597–1606. https://doi.org/10.1093/jac/dks121 PMid:22499996

Clinical Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing, 25 thin formational supplements. CLSI, document M100-S25, Wayne PA, 2015.

Nordmann P, Gniadkowski M, Giske CG, et al. Identification and screening of carbapenemase-producing Enterobacteriaceae. Clinical Microbiology and Infection. 2012; 18 (5): 432–438. https://doi.org/10.1111/j.1469-0691.2012.03815.x PMid:22507110

Patel J, Cockerill F, Bradford P, et al. Clinical and Laboratory Standards Institute, Performance Standards for Antimicrobial Susceptibility Testing; Twenty first Informational Supplement. CLSI document M100-21. Wayne, PA: Clinical and Laboratory Standards Institute, 2011: 35(3).

Nordmann P, Poirel L. Strategies for identification of carbapenemase-producing Enterobacteriaceae. J Antimicrob Chemother. 2013; 68:487–489. https://doi.org/10.1093/jac/dks426 PMid:23104494

Huang T, Poirel L, Pierre, Bogaerts P et al. Temocillin and piperacillin/tazobactam resistance by disk diffusionas antimicrobial surrogate markers for the detection of carbapenemase-producing Enterobacteriaceae in geographical areas with a high prevalence of OXA-48 producers. J Antimicrob Chemother. 2014; 69: 445–450. https://doi.org/10.1093/jac/dkt367 PMid:24055766

Monteiro J, Widen H R, Pignatari C A, et al. Rapid detection of carbapenemase genes by multiplex real-time PCR. Journal of Antimicrobial Chemotherapy. 2012; 67(4), 906-909. https://doi.org/10.1093/jac/dkr563 PMid:22232516

Sanjeev K, Mehra SK. Performance of Modified Hodge Test and Combined Disk Test for Detection of Carbapenemases in Clinical Isolates of Enterobacteriaceae. Int J Curr Microbiol App Sci. 2015: 772-783.

Glupczynski Y, Huang TD, Bouchahrouf W, et al. Rapid emergence and spread of OXA-48-producing carbapenem-resistant Enterobacteriaceae isolates in Belgian hospitals. Int J Antimicrob Agents. 2012; 39:168 –172. https://doi.org/10.1016/j.ijantimicag.2011.10.005 PMid:22115539

Hartl R, Widhalm S, Kerschner H, et al. Temocillin and meropenem to diskriminate resistance mechanisms leading to decreased carbapenem susceptibility with focus on OXA-48 in Enterobacteriaceae. Clin Microbiol Infect. 2013; 19: E230–232. https://doi.org/10.1111/1469-0691.12146 PMid:23397897

Doyal D, Peirano G, Lascols C, et al. Laboratory detection of Enterobacteriaceae that produce carbpenemases. J, Clin. Microbiol. 2012; 50(12):3877-3880. https://doi.org/10.1128/JCM.02117-12 PMid:22993175 PMCid:PMC3503014

Andrea B, Ilaria F, Antonietta C, et al. Comparison of phenotypic methods for the detection of carbapenem non-susceptible Enterobacteriaceae. J BMC Infectious Diseases. 2014; 6:3-7.

Canton R, Akova M, Carmeli Y, et al. Rapid evolution and spread of carbapenemases among Enterobacteriaceae in Europe. Clinical Microbiology and Infection. 2012; 18(5) 414-431. https://doi.org/10.1111/j.1469-0691.2012.03821.x PMid:22507109

UK Standards for Microbiology Investigations (UK SMI). Detection of bacteria with carbapenem-hydrolysing β-lactamases (carbapenemases), PHE Bacteriology, 2015: 60.

Samar S. Comparative evaluation of phenotypic and genotypic methods for detection of carbapenemases in clinically significant Klebsiella pneumonia isolates. Egyptian Journal of Medical Microbiology. 2016; 25(1):109-116. https://doi.org/10.12816/0037099

Woodford N, Pike R, Meunier D, et al. In vitro activity of temocillin against multidrug-resistant clinical isolates of Escherichia coli, Klebsiella spp. And Enterobacter spp., and evaluation of high-level temocillin resistance as a diagnostic marker for OXA-48 carbapenemase. J Antimicrob. Chemother. 2014; 69:564-567. https://doi.org/10.1093/jac/dkt383 PMid:24080500

Livermore DM, Warner M, Mushtaq S et al. What remains against carbapenem-resistant Enterobacteriaceae? Evaluation of chloramphenicol, ciprofloxacin, colistin, fosfomycin, minocycline, nitrofurantoin, temocillin and tigecycline. Int J Antimicrob Agents. 2011; 37: 415–419. https://doi.org/10.1016/j.ijantimicag.2011.01.012 PMid:21429716

Published

2018-02-01

How to Cite

1.
Shaker OA, Gomaa HEED, ElMasry SA, Abdel Halim RM, Abdelrahman AH, Kamal JS. Evaluation of Combined Use of Temocillin Disk and Mastdisks Inhibitor Combination Set Against Polymerase Chain Reaction for Detection of Carbapenem-Resistant Enterobacteriaceae. Open Access Maced J Med Sci [Internet]. 2018 Feb. 1 [cited 2024 Nov. 4];6(2):242-7. Available from: https://oamjms.eu/index.php/mjms/article/view/oamjms.2018.090

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Section

A - Basic Science