Introduction: Infections caused by multidrug-resistant Klebsiella pneumoniae are difficult to treat and pose a serious threat to public health worldwide. Here, we describe the presence of carbapenemase-producing K. pneumoniae in intensive care units (ICU) of three major Mato Grosso do Sul hospitals located in the Midwest region of Brazil. Methodology: A total of 165 K. pneumoniae isolates with reduced susceptibility to carbapenems as identified by the VITEK-2 compact system were studied. Antimicrobial susceptibility testing was performed using the disk diffusion method, as recommended by the Clinical and Laboratory Standards Institute, and the E-test method. The detection of carbapenemase was performed using the modified Hodge test and polymerase chain reaction. Results: The blaKPC gene was identified in 88.1% (n=89) of the selected K. pneumoniae isolates from Beneficent Association of Campo Grande, 94.9% (n=34) of the isolates from the Regional Hospital of Mato Grosso do Sul and 95.2% (n=26) of the isolates from Maria Aparecida Pedrossian University Hospital. Resistance greater than 80% was observed against cephalosporins, aztreonam, ciprofloxacin and piperacillin/tazobactam. Carbapenemase-producing K. pneumoniae (Kp-KPC) isolates were considered important causative agents of urinary tract infections, pneumonia and bloodstream infections in ICU patients. While rarely reported in the literature, we documented three cases of meningoencephalitis caused by Kp-KPC. Conclusions: Our study documents the presence of Kp-KPC in three major Mato Grosso do Sul state hospitals, providing key national epidemiology data. This is an important mechanism of resistance in K. pneumoniae isolates from ICU patients and is associated with resistance to multiple classes of antimicrobial drugs.