Phenotypic and Molecular Characterization of Extended Spectrum Beta-lactamase Producing Escherichia Coli and Klebsiella Pneumoniae in Clinical Isolates at Embu Level Five Hospital and Kenyatta National Hospital, Kenya.

Abstract

Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) are considered by the World Health Organization to be the predominant priority human bacterial pathogens. The pathogens cause blood stream infections, urinary tract infections, and hospital-acquired pneumonia. The production of extended spectrum β-lactamases (ESBLs) among those two pathogens have contributed to the occurrence and transmission of antimicrobial resistance against beta lactam antibiotics and various other categories of antimicrobial agents in hospital settings. These multi drug resistant (MDR) bacterial infections have led to limited therapeutic options, increased healthcare costs and mortalities. Therefore, this study established ESBL producing and multidrug resistant E. coli and K. pneumoniae isolated from patients in Embu Level 5 and Kenyatta National Hospitals, Kenya. The specific objectives were; (i) To identify the E. coli and K. pneumoniae infecting patients treated at Embu Level 5 Hospital and Kenyatta National Hospital (ii) To determine the phenotypic antimicrobial resistance profiles of E. coli and K. pneumoniae infecting patients treated at Embu Level 5 Hospital and Kenyatta National Hospital (iii) To establish the genetic determinants responsible for the resistance phenotypes of E. coli and K. pneumoniae including MDR isolates. A cross-sectional laboratory-based study design was adopted whereby 138 E. coli and 127 K. pneumoniae samples were collected from various clinical specimens at the two health facilities from January 2020 to Feb 2021. The isolates were analyzed at the Pharmacology, Public Health and Toxicology Laboratory, UoN. Phenotypic confirmatory disk diffusion tests for ESBLs and antibiotic susceptibility testing were done according to the Clinical Laboratory Standards Institute guidelines (2020). Molecular analysis was done through conventional Polymerase Chain Reaction (PCR) utilizing primers for gadA, rpoB and selected ESBL genes. Forty-two representative samples were taken for sequencing and ...