Evaluation of the compatibility of phenotypic and molecular methods used to determine carbapenem resistance in enterobacterales isolates

Aim: Carbapenems are one of the most important options for clinicians with few treatment options in the clinic due to their low sideeffects, rapid diffusion into tissues and use in all age groups. Therefore, it is important to be able to detect carbapenemase-producingisolates at an early stage for appropriate patient management and for infection prevention and control procedures. Antibioticresistance genes and enzymes of microorganisms can be determined by phenotypic and molecular methods in clinical microbiologylaboratories. Phenotypic methods are cheap, easy and easy to repeat but determination of resistance gene regions by molecularmethods is costly, requires labour-experienced personnel and is time consuming. Determining whether the isolates possess thecarbapenemase enzyme by phenotypic tests will provide convenience both for the patient and early initiation of the treatment anddirecting the clinician to the treatment. In this study, we aimed to evaluate the compatibility of phenotypic methods (carbapenemaseinactivation method and Rapidec Carba NP) and molecular methods (Polymerase Chain Reaction) used to determine carbapenemresistance in Enterobacterales isolates.Material and Methods: Carbapenem resistant 60 and sensitive 20 Enterobacterales isolates were included in the study. E-test agargradient diffusion, CIM, Rapidec Carba NP methods and PCR were studied. The agreement between the methods was determined byusing the kappa (κ) coefficient with the cohen kappa analysis method.Results: In carbapenem resistant isolates, meropenem MİK50 and MİK90 determined as 32µg/ml, 64µg/ml, imipenem MİK50 andMİK90 determined as 32µg/ml, 128µg/ml, respectively. OXA-48 was positive in 54 (90%) isolates and NDM-1 in 6 (10%) isolates.The susceptibility of the isolates with OXA-48 carbapenemase gene region was 94.4% by CIM test and 92.6% by Rapidec Carba NPtest, respectively. When the Kappa coefficient was evaluated, a very good agreement was observed between both tests and OXA-48.However, in the isolates with NDM-1 gene region, no compliance with CIM test was observed but Rapidec Carba NP test showedvery good agreement.Conclusion: Rapid carbapenemase testing, such as Rapidec Carba NP and CIM, can play an important role in preventing thedevelopment of health-related outbreaks caused by carbapenemase-producing isolates, enabling faster prevention and control ofinfection.

PDF

___

1. Van Duin D, Doi Y. The global epidemiology of carbapenemase-producing Enterobacteriaceae. Virulence 2017;8:460-9.
2. Djahmi N, Dunyach-Remy C, Pantel A, et al. Epidemiology of Carbapenemase-Producing Enterobacteriaceae and Acinetobacter baumannii in Mediterranean Countries. Biomed Res Int 2014;2014:305784.
3. Nørgaard SM, Jensen CS, Aalestrup J, et al. Choice of therapeutic interventions and outcomes for the treatment of infections caused by multidrug-resistant gram-negative pathogens: a systematic review. Antimicrob Resist Infect Control 2019;4:170.
4. Duman Y, Ersoy Y, Gursoy NC, et al. A silent outbreak due to Klebsiella pneumoniae that co-produced NDM1 and OXA-48 carbapenemases, and infection control measures. Iran J Basic Med Sci 2020;23:46-50.
5. US Centers for Disease Control and Prevention. Antibiotic/antimicrobial resistance: biggest threats. Atlanta (GA): CDC;2013.
6. Duman Y, Kuzucu C, Tekerekoglu MS, et al. Changing trends of carbapenem resistance of Escherichia coli and Klebsiella pneumoniae strains isolated from intensive care units, inpatient services and outpatient’s clinics: a five years’ retrospective analysis. Med Science 2018;7:536-9.
7. Hrabak J, Študentová V, Walková R, et al. Detection of NDM-1, VIM-1, KPC, OXA-48, and OXA-162 Carbapenemases by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry. J Clin Microbiol 2012;50:2441-3.
8. Poirel L, Walsh TR, Cuvillier V, et al. Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis 2011;70:119–23.
9. Alpar R. Spor, Sağlık ve Eğitim Bilimlerinden Örneklerle Uygulamalı İstatistik ve Geçerlik-Güvenirlik. 1. baskı. Detay Yayıncılık, Ankara, 2010; 20-45.
10. Tao XB, Qian LH, Li Y, et al. Hospital acquired infection rate in a tertiary care teaching hospital in China: a cross-sectional survey involving 2434 in patients. Int J Infect Dis 2014;27:7-9.
11. Gijon D, Curiao T, Baquero F, et al. Fecal carriage of carbapenemase-producing Enterobacteriaceae: a hidden reservoir in hospitalized and non-hospitalized patients. J Clin Microbiol 2012;50:1558–63.
12. Hauck C, Cober E, Richter SS, et al. Spectrum of excess mortality due to carbapenem-resistant Klebsiella pneumoniae infections. Clin Microbiol Infect 2016;22:513–9.
13. Nordmann P, Poirel L. The difficult-to-control spread of carbapenemase producers among Enterobacteriaceae worldwide. Clinical Microbiology and Infection 2014;20:821–30.
14. Giske CG, Martinez L, Cantón R, et al. EUCAST guidelines for detection of resistance mechanisms and specific resistances of clinical and / or epidemiological importance. Version 2.0, EUCAST, Switzerland, 2017:4-30.
15. Kabir MH, Meunier D, Hopkins KL, et al. A two-centre evaluation of rapidec carba np for carbapenemase detection in Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter spp. J Antimicrob Chemother 2016;71:1213-6.
16. Davarcı İ, Koçoğlu ME, Zengin F. Comparison of carbapenem inactivation test and modified Hodge test in carbapenemase-producing Klebsiella pneumoniae strains. Klimik Derg 2018;31:223-6.
17. Bayramoğlu G, Uluçam G, Gençoğlu ÖÇ, et al. Comparison of the modified Hodge test and the Carba NP test for detection of carbapenemases in Enterobacteriaceae isolates. Mikrobiyol Bul 2016;50:1-10.
18. Hombach M, von Gunten B, Castelberg C, et al. Evaluation of the Rapidec Carba NP test for detection of carbapenemases in Enterobacteriaceae. J Clin Microbiol 2015;53:3828–33.
19. Aktaş E, Malkoçoğlu G, Otlu B, et al. Evaluation of the Carbapenem Inactivation Method for Detection of Carbapenemase-Producing Gram-Negative Bacteria in Comparison with the Rapidec Carba NP. Microb Drug Resist 2017;23:457-461.
20. Tijet N, Patel SN, Melano RG. Detection of carbapenemase activity in Enterobacteriaceae: comparison of the carbapenem inactivation method versus the Carba NP test. J Antimicrob Chemother 2016;71:274-6.
21. Yamada K, Kashiwa M, Arai K, et al. Comparison of the Modified- Hodge test, Carba NP test, and carbapenem inactivation method as screening methods for carbapenemase-producing Enterobacteriaceae. J Microbiol Methods 2016;128:48–51.
22. Osterblad M, Hakanen AJ, Jalava J. Evaluation of the Carba NP test for carbapenemase detection. Antimicrob Agents Chemother 2014;58:7553-6.
23. Dortet L, Bréchard L, Cuzon G, et al. Strategy for rapid detection of carbapenemase-producing Enterobacteriaceae. Antimicrob Agents Chemother 2014;58:2441-5.