What type of study is this?

This is a Singleplex PCR study (also classified as: Multiplex PCR).

September 10, 2025PLoS ONE0 citationsOpen Access

Carbapenemase-encoding genes in critical gram-negative bacteria isolated from ICU patients with infections and/or gastrointestinal carriage, and environmental samples in the amhara National Regional state, Ethiopia

MKMizan KinduMadda Walabu University FMFeleke MogesUniversity of Gondar AAAshenafi AlemuArmauer Hansen Research Institute

Key Points

Carbapenem-resistant isolates showed diverse encoding genes, indicating significant infection risks in ICU settings.
The predominant gene, bla NDM, was detected in 75.9% of carbapenem-resistant isolates, showcasing a critical public health concern.
Singleplex and multiplex PCR were used to analyze the presence of carbapenemase genes in various sources, reinforcing the study's robust methodology.
The presence of multiple carbapenemase genes in a notable proportion of isolates emphasizes the urgent need for effective infection control measures.

Abstract

Effective infection control requires identifying and eliminating carbapenemase-producing (CP) Gram-negative bacteria (GNB) in high-risk groups like intensive care unit (ICU) patients and from contaminated environmental surfaces. This study aimed to describe the diversity of carbapenemase-encoding genes among critical GNB isolates from ICU patients with infection and/or gastrointestinal (GI) colonization, as well as from ICU environmental surfaces in the Amhara National Regional state, Ethiopia.A total of 169 carbapenem-resistant isolates were identified, including 26 from infections, 82 from GI colonization, and 61 from environmental samples. These comprised Klebsiellapneumoniae (n = 107), Escherichia coli (n = 16), Pseudomonas aeruginosa (n = 9), and Acinetobacter species (n = 37), of which 147 were analyzed.Singleplex and multiplex PCR were performed to detect predominant carbapenemase-encoding genes, including KPC ( bla KPC ), MBLs ( bla IMP , bla VIM , and bla NDM ), and OXA ( bla OXA-48 , bla OXA-23 , and bla OXA-58 ). PCR analysis revealed that at least one carbapenemase-encoding gene was detected in 133 (78.7%) of the 169 carbapenem-resistant isolates from patients and environmental surfaces. They were detected in 22/133(16.5%), 70/133(52.6%), and 41/133(30.8%) isolates obtained from infection, GI colonization, and environmental samples, respectively. The most prevalent carbapenemase-encoding gene, bla NDM , was found in 101 (75.9%) of the 133 CP isolates. Other detected carbapenemase-encoding genes included bla OXA-23 in 5 (3.8%) isolates, bla VIM in 4 (3.0%), bla OXA-48 in 1 (0.8%), and bla OXA-58 in 1 (0.8%). K. pneumoniae and Acinetobacter spp. harbored all the gene types detected in this study. Co-harboring of two or more carbapenemase-encoding genes, in combination with bla NDM , was observed in 21 of 133 (15.8%) isolates, including 7 K. pneumoniae and 14 Acinetobacter spp. isolates. However, bla KPC and bla IMP were not identified in any of the tested isolates. This study highlights the presence of carbapenemase-encoding genes among critical GNB across all sample source types in the study area. Moreover, the detection of isolates harboring multiple carbapenemase-encoding genes underscores the need for enhanced infection control measures in ICU settings.

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