Genetic Evaluation of Heteroresistance among Carbapenem-Susceptible Clinical Isolates of Enterobacterales

Abstract

Carbapenems currently serve as the last line of defense when treating serious infections caused by multidrug-resistant Enterobacterales species; however, heteroresistance of these species is thought to cause failure in the treatment with these broad-spectrum antibiotics. This study was designed to determine the prevalence of carbapenem heteroresistance and associated genotypic modifications among phenotypically meropenem-susceptible Escherichia coli and Klebsiella pneumoniae isolates. A total of 204 isolates of E. coli (n: 118) and K. pneumoniae (n: 86) from various clinical samples were included in this prospective experimental study. Identification and antimicrobial susceptibility testing of the isolates were performed by VITEK® (bioMérieux, France). Strains that were found susceptible to carbapenem group antibiotics (meropenem, imipenem, and ertapenem) with automated system were further investigated by disk diffusion method. The isolates with discrete colony growth within the clear inhibition zone among phenotypically meropenem-susceptible strains were tested for heteroresistance with the “gold standard” population analysis profile-area under the curve (PAP-AUC) method. In addition, heteroresistant isolates were analyzed for the presence of carbapenemase genes with in-house PCR method. The heteroresistance prevalence rate was 3.5% for E. coli and 18.1% for K. pneumoniae. The presence of heteroresistance in a total of 10 meropenem-susceptible isolates (E. coli, n: 4; K. pneumoniae, n: 6) was confirmed by the PAP-AUC method. The most frequently detected carbapenemase in heteroresistant isolates was OXA-48 (6/10), followed by NDM-1 (2/10). Meropenem is frequently preferred as initial empirical monotherapy in most of Gram-negative infections in adult and pediatric patients. The presence of heteroresistance against meropenem is too important to ignore, and for this reason, it seems beneficial to prefer combined treatment regimens in clinical practice.