Phenotypic and Genotypic Evaluation of Carbapenamase Producing Klebsiella Pneumoniae Isolates with Their Phylogenetic Analysis at an Egyptian University Hospital

Abstract

Widespread dissemination of carbapenem-producing Klebsiella pneumoniae (KPC) is of major concern in healthcare settings. Resistance to carbapenems involves multiple mechanisms such as the production of carbapenemases, impermeability of outer membrane and efflux pump mechanism. Objective: The aim of this study was to evaluate the prevalence of carbapenemase-producing K. pneumoniae strains among various clinical specimens obtained from different wards and to detect KPC as a mechanism of resistance. Methodology: 100 samples (55urine and 45sputum) were collected from outpatients and inpatients attending urology and chest departments in Beni Suef University Hospital aiming to isolate K. pneumoniae during the period of December 2016 to January 2018. The isolates were tested for susceptibility to ertapenem using E test. Resistant isolates were subjected to phenotypic detection of carbapenemase production by Modified Hodge Test (MHT) and molecular assessment of KPC gene by PCR. Phylogentic tree analysis was used to detect their relationship by DNA sequencing reaction. Results: K.pneumoniae were isolated from 31(31%) of the samples taken. Out of them 19(61.8%) were resistant to ertapenem by E test. By phenotypic method,17/19 (89.4%) were positive for carbapenemase by MHT; and only 13 out of them (76.4%) were confirmed as KPC by PCR. Conclusion: High rate of carbapenem-resistance in K. pneumoniae by both phenotypic and molecular methods was observed. These results warrant more firm infection control measures along with a strictly implemented antibiotic stewardship program to prevent their spread.