Detection and molecular characterization of carbapenem-resistant gram-negative bacterial isolates

Abstract

Antimicrobial-resistant bacteria (ARB) are responsible for increased mortality and morbidity. Therefore, this study focuses on evaluating traditional and molecular diagnostic tools of carbapenem-resistant gram-negative bacteria (CRGNB). In order to achieve this, 94 samples, from different patients' specimens, and surrounding environment, were collected from intensive care units (ICUs) at Ain Shams University Specialized Hospital and the National Cancer Institute, Cairo, Egypt. The swabs were cultured on appropriate media, including Chromogenic medium (HiCrome KPC Agar Base "HIMEDI AM1831"), and MacConkey-10 µg imipenem disc resulting in 136 isolates with different culture characteristics. Next, the selected isolates were subjected to VITEK 2 machine and 16SrRNA (16 S ribosomal RNA) sequencing. The sensitivity of HiCROME KPC agar for CRGNB detection was 99.3% and 94.7%, in reference to the MacConkey-disc and VITEK-2 methods, respectively. The HiCrome KPC agar assumptions for bacterial identification were not as consistent as those of VITEK 2 (with only 47.4% agreement) and 16SrRNA gene sequencing analysis. The approaches discussed in this study facilitate providing rapid diagnosis and treatment of CRGNB, which helps increase survival rates. HiCrome KPC agar is considered a relatively accurate and easy method that can be used in any laboratory. In addition, the selected strains were deposited in the gene bank with the accession numbers OR553657, OR553658, and OR553659. It is noteworthy that Genus Acinetobacter is the major CRGNB isolated from the patients and environmental surfaces in the hospitals. This highlights the importance of proper environmental and terminal cleaning procedures in healthcare facilities and applying control measures to ensure infection prevention.