In Vitro Activity of Single and Combined Antibiotics against Carbapenem Resistant Enterobacteriaceae Clinical Isolates in Relation to their Resistance Genes

Inas El-Defrawy; Aisha Abu Aitta; Nevine Fam; Manar Khaled; Nadia Madany; Mervat El Damarawy; Doaa Gamal; Mohammed Amr Alkholy

doi:10.3889/oamjms.2022.10347

Authors

Inas El-Defrawy Department of Microbiology, Theodor Bilharz Research Institute, Giza, Egypt
Aisha Abu Aitta Department of Microbiology, Theodor Bilharz Research Institute, Giza, Egypt https://orcid.org/0000-0002-6375-9618
Nevine Fam Department of Microbiology, Theodor Bilharz Research Institute, Giza, Egypt
Manar Khaled Department of Microbiology, Theodor Bilharz Research Institute, Giza, Egypt https://orcid.org/0000-0002-0674-8031
Nadia Madany Department of Medical Microbiology and Immunology, Faculty of Medicine, Cairo University, Giza, Egypt
Mervat El Damarawy Intensive Care Unit, Theodor Bilharz Research Institute, Giza, Egypt
Doaa Gamal Department of Microbiology, Theodor Bilharz Research Institute, Giza, Egypt https://orcid.org/0000-0002-6279-8105
Mohammed Amr Alkholy Department of Medical Microbiology and Immunology, Faculty of Medicine, Cairo University, Giza, Egypt

DOI:

https://doi.org/10.3889/oamjms.2022.10347

Keywords:

Carbapenem, Treatment, Carbapenem-resistant Enterobacteriaceae, Carbapenemase-producing Enterobacteriaceae, Colistin, Tygecycline, Dual carbapenem, NDM, OXA-48

Abstract

Background: Mortality due to infection with carbapenem-resistant Enterobacteriaceae (CRE) is reported globally and carbapenemase production is the main mechanism of resistance in these isolates. The detection and treatment of carbapenemase-producing Enterobacteriaceae (CPE) is a major challenge in health care facilities.

Objectives: The aim of the current study was to evaluate the in-vitro effect of different single and combined antibiotic agents against CRE clinical isolates.

Methodology: Fifty CRE isolates were detected using disk diffusion test as a screening test. Species identification and antibiotic susceptibility testing was done using Vitek 2 system. Carbapenemase enzyme production was confirmed by Carba NP test. Multiplex PCR was done to detect carbapenem resistance genes. Antibiotics were tested in the form of single agents (colistin and tigecycline) and combined (tigecycline/ colistin, doripenem/ colistin and dual carbapenem therapy (ertapenem and doripenem) against CRE isolates using E-test method.

Results: Most of the CRE isolates were K. pneumoniae, 68%, followed by E. coli, 22%, S. marcescens, 4%, E. cloacae, 4% and C. freundii, 2%. CPE was confirmed in 46 isolates by multiplex PCR; bla_NDM-like was the main carbapenem resistance gene in (84%) of the isolates, followed by bla_OXA-48-like (6%) and bla_KPC-like (2%). Carba NP test detected 90% of CPE isolates. Single use of colistin and tigecycline showed 100% sensitivity against all tested CRE isolates except in bla_NDM-like (83%). Combination of colistin/tigecycline showed synergetic activity in 18% of CRE that was correlated to their carbapenemase R genes showing a significant increase in bla_OXA-48-like and bla_KPC-like positive isolates (100%) compared to bla_NDM-like (7%). Other combinations showed indifferent effect whereas antagonism was not detected in any of the tested combinations.

Conclusions: bla_NDM-like is the main carbapenemase-producing gene detected among our CPE isolates followed by bla_OXA-48-like. Colistin and tigecycline are still effective when used as single agents, and may offer effective treatment options when used in combination for CRE infections. Characterization of carbapenemases is crucial in determining treatment options. There is urgent demand for the development of novel therapeutic agents against NDM-producing CPE isolates.

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