Rapid screening for carbapenemase-producing carbapenem-resistant <i>Enterobacterales</i> : clinical implementation of an immunochromatographic test using broth-enriched rectal swabs
Camila Loredana Pereira Alves Madeira Bezerra, Barbara de Almeida Lessa Castro, Anna S. Levin, Lindissy Luara Baldi, Ana Paula Cury, Flávia Rossi, Natashia Reese, Fernanda C. Lessa, et al. (35 authors)
Microbiology Spectrum · 2025-08
Abstract
ABSTRACT Carbapenemase-producing carbapenem-resistant Enterobacterales (CP-CRE) poses a public health issue. Rapid detection of CP-CRE colonization is challenging; existing methods are either expensive or time-consuming. We evaluated an immunochromatographic test (ICT) for detecting carbapenemases directly from broth-enriched rectal swabs. One hundred intensive care patients provided 178 pairs of rectal swabs. One swab was tested using the GeneXpert Carba-R PCR assay; the other was inoculated into brain-heart infusion broth. After 4 and 6 h of incubation at 37°C, the broth was tested with the RESIST-5 O.K.N.V.I. ICT for Klebsiella pneumoniae carbapenemase (KPC), New Delhi metallo-β-lactamase (NDM), oxacillinase-48 (OXA-48), Verona integron-encoded metallo-β-lactamase (VIM), and imipenemase (IMP) carbapenemases. Broths were subcultured after overnight incubation, and recovered carbapenem-resistant Enterobacterales isolates were tested using GeneXpert Carba-R PCR. Sensitivity, specificity, and accuracy were calculated in comparison to culture positive for CP-CRE, confirmed by PCR for KPC, NDM, OXA-48, VIM, and IMP. Of the 178 swabs, 60 were culture positive for CP-CRE. After 6 h, the ICT demonstrated a sensitivity of 65%, specificity of 97.5%, and accuracy of 86.8%. Among heavily soiled swabs, sensitivity reached 81.8% for ICT after 6 h, and the specificity was 100%. The mean execution time for carbapenemase detection using ICT was reduced by 60 h compared to culture. The ICT after 6 h incubation offers reduced execution time for detecting CP-CREs. This method may serve as a valuable rapid screening tool, especially in resource-limited settings. IMPORTANCE The rapid spread of multidrug-resistant bacteria requires innovative solutions for early detection and prevention measures. In this study, we present a simple protocol for the direct detection of carbapenemases in rectal swabs using an immunochromatographic assay. By optimizing the assay conditions, we achieved rapid and high-accuracy identification of five clinically important carbapenemases. This method can broaden access to rapid CP-CRE detection of fecal colonization—even in laboratories with limited resources—enabling the implementation of faster and more effective infection control measures, potentially reducing the spread of resistance.
MeSH terms
- GeneXpert MTB/RIF
- Klebsiella pneumoniae
- Microbiology
- Incubation
- Carbapenem
- Medicine
- Incubation period
- Biology
- Virology