Rapid screening for carbapenemase-producing carbapenem-resistant <i>Enterobacterales</i> : clinical implementation of an immunochromatographic test using broth-enriched rectal swabs
Carbapenemase-producing carbapenem-resistant <i>Enterobacterales</i> (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 <i>Klebsiella pneumoniae</i> 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 <i>Enterobacterales</i> 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.IMPORTANCEThe 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.
Authors
- Sílvia Figueiredo Costa (ORCID: https://orcid.org/0000-0003-1087-752X)
- Chung-Chou H. Chang (ORCID: https://orcid.org/0000-0001-8719-2508)
- Roberta Muriel Longo Roepke (ORCID: https://orcid.org/0000-0003-2214-5166)
- Pedro Fortes Osório Bustamante (ORCID: https://orcid.org/0000-0001-9383-4722)
- María De Jesús (ORCID: https://orcid.org/0000-0002-6757-2746)
- Susan Bollinger
- Thaís Guimarães (ORCID: https://orcid.org/0000-0002-7282-5453)
- Maristela Pinheiro Freire (ORCID: https://orcid.org/0000-0002-9691-192X)
- Matias Chiarastelli Salomão (ORCID: https://orcid.org/0000-0001-5507-0063)
- Estêvão Bassi (ORCID: https://orcid.org/0000-0002-3819-2699)
- Leandro Utino Taniguchi (ORCID: https://orcid.org/0000-0003-4384-0408)
- Anna S. Levin (ORCID: https://orcid.org/0000-0003-2427-8368)
- Ana Natiele da Silva Barros
- Luíz Marcelo Sá Malbouisson (ORCID: https://orcid.org/0000-0002-3261-5603)
- Fernanda Spadão
- Fernanda C. Lessa (ORCID: https://orcid.org/0000-0002-8031-6376)
- Barbara de Almeida Lessa Castro
- A Teruel
- Ludhmila Abrahão Hajjar (ORCID: https://orcid.org/0000-0001-5645-2055)
- Flávia Rossi (ORCID: https://orcid.org/0000-0002-6243-6606)
- Tania C. T. Ferraz Alves (ORCID: https://orcid.org/0000-0003-3402-9656)
- Natashia Reese
- Priscila Koba Kodato
- Ícaro Boszczowski (ORCID: https://orcid.org/0000-0002-0221-3814)
- Laina Bubach
- Catarina Dinis Fernandes (ORCID: https://orcid.org/0000-0002-3967-5836)
- Marcelo Park
- Ana Paula Cury (ORCID: https://orcid.org/0000-0002-4820-9781)
- Camila Loredana Pereira Alves Madeira Bezerra
- Alexandre Pereira Funari
- Lindissy Luara Baldi
- Julia Fontanesi
- Ana Clara Dobre
- Edivaldo Utyiama
Institutions
- Centers for Disease Control and Prevention (US)
- Universidade de São Paulo (BR)
- Centers for Disease Control and Prevention (UG)
- Instituto de Medicina Tropical (PY)
- Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (BR)
- Fleury S.A. (Brazil) (BR)
- Fundação Faculdade de Medicina (BR)
Publication Details
- Journal
- Microbiology Spectrum
- Published
- 2025-08-08
- DOI
- https://doi.org/10.1128/spectrum.01313-25
- Primary Topic
- Antibiotic Resistance in Bacteria
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Fundação de Amparo à Pesquisa do Estado de São Paulo
- Centers for Disease Control and Prevention