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.

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Journal
Microbiology Spectrum
Published
2025-08-08
DOI
https://doi.org/10.1128/spectrum.01313-25
Primary Topic
Antibiotic Resistance in Bacteria
Type
article
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article

Rapid screening for carbapenemase-producing carbapenem-resistant Enterobacterales : clinical implementation of an immunochromatographic test using broth-enriched rectal swabs

Sílvia Figueiredo Costa, Chung-Chou H. Chang, Roberta Muriel Longo Roepke, Pedro Fortes Osório Bustamante et al.
Microbiology Spectrum
Antibiotic Resistance in Bacteria
article

Rapid screening for carbapenemase-producing carbapenem-resistant Enterobacterales : clinical implementation of an immunochromatographic test using broth-enriched rectal swabs

Sílvia Figueiredo Costa, Chung-Chou H. Chang, Roberta Muriel Longo Roepke, Pedro Fortes Osório Bustamante, María De Jesús, Susan Bollinger, Thaís Guimarães, Maristela Pinheiro Freire, Matias Chiarastelli Salomão, Estêvão Bassi, Leandro Utino Taniguchi, Anna S. Levin, Ana Natiele da Silva Barros, Luíz Marcelo Sá Malbouisson, Fernanda Spadão, Fernanda C. Lessa, Barbara de Almeida Lessa Castro, A Teruel, Ludhmila Abrahão Hajjar, Flávia Rossi, Tania C. T. Ferraz Alves, Natashia Reese, Priscila Koba Kodato, Ícaro Boszczowski, Laina Bubach, Catarina Dinis Fernandes, Marcelo Park, Ana Paula Cury, Camila Loredana Pereira Alves Madeira Bezerra, Alexandre Pereira Funari, Lindissy Luara Baldi, Julia Fontanesi, Ana Clara Dobre, Edivaldo Utyiama
article en

Abstract

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.

Microbiology SpectrumVol. 13(9)
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)
Fundação de Amparo à Pesquisa do Estado de São Paulo, Centers for Disease Control and Prevention
Openalex Percentile: Top 84%
Antibiotic Resistance in Bacteria
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