Development of simplex and multiplex immunochromatographic assays for rapid detection of GES-, PER-, and VEB-producing gram-negative bacteria
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Le résumé fourni par la source
ABSTRACT Extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae are increasingly reported worldwide, emphasizing the need for rapid diagnostic tools to limit their spread and prevent outbreaks. Although CTX-M enzymes remain predominant, less common ESBLs such as GES, PER, and VEB are emerging and gaining clinical relevance. We developed simplex and multiplex lateral flow immunoassays (LFIAs) for the rapid detection of these ESBLs. Monoclonal antibodies were produced and incorporated into dedicated LFIA strips. The assays were evaluated using 116 WGS-characterized gram-negative bacteria (GNB) isolates expressing diverse β-lactamases and grown on various media. For testing, three colonies were suspended in 150 µL of extraction buffer, applied to the strips, and results were visually interpreted after 15 min. Both simplex and multiplex LFIAs achieved 100% sensitivity and specificity for detecting GES-, PER-, and VEB-producing isolates, irrespective of bacterial species, β-lactamase content, or growth conditions. The panel included prevalent variants of PER ( n = 5, including PER-2), VEB ( n = 9), and GES ( n = 9, including ESBLs and carbapenemases). Detection limits for recombinant PER-1, VEB-1, and GES-1 were 5, 50, and 100 pg/mL, respectively, and 10 5 , 10 6 , and 10 6 CFU/mL for corresponding Acinetobacter baumannii producers grown in tryptic soy broth. Overall, these simplex and multiplex GES-, PER-, and VEB-targeting LFIAs provide a rapid, accurate, and user-friendly approach suitable for routine use in clinical microbiology laboratories. When combined with the NG-Test CTX-M MULTI assay, which detects the five major CTX-M groups, they may further enhance the overall detection of the most common ESBLs, particularly among non-fermenting GNB. IMPORTANCE Although less prevalent globally than CTX-M-type ESBLs, GES-, PER-, and VEB-type β-lactamases represent an important and often underrecognized mechanism of resistance, particularly in non-fermenting gram-negative pathogens. These enzymes are frequently associated with multidrug-resistant organisms and may significantly compromise available therapeutic options. The detection of the genes encoding these enzymes is often based on homemade PCR. Rapid diagnostic tests based on lateral flow immunoassays (LFIAs) have been successfully used to detect CTX-M-like ESBLs and the five main carbapenemases. Here, we demonstrated that GES, PER, and VEB enzymes can be readily detected by simplex or multiplex LFIAs from Enterobacterales, Pseudomonas aeruginosa , and Acinetobacter baumannii colonies. These assays are rapid (≤15 min), highly sensitive (10 5 –10 6 CFU/mL), requiring no specialized equipment, thus offering a user-friendly alternative to molecular assays in clinical laboratories. These features align well with the World Health Organization ASSURED criteria (Affordable, Sensitive, Specific, User-friendly, Rapid, Equipment-free, and Deliverable).
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Le contrôle bibliographique ouvert
DOI retrouvé dans Crossref DOI retrouvé ; titre concordant.
- Titre Crossref
- Development of simplex and multiplex immunochromatographic assays for rapid detection of GES-, PER-, and VEB-producing gram-negative bacteria
- Date Crossref
- 11/09/2026
- Éditeur
- American Society for Microbiology
- Type
- journal-article
Ce recoupement confirme des métadonnées liées au DOI. Il ne confirme ni la méthode ni les conclusions de l’étude, et il ne compte pas comme une seconde source scientifique indépendante.
Où se fait cette recherche
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University of Sarajevo Department of Pathobiology and Epidemiology pays non établi dans la noticeUniversité ou école supérieure
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Inserm pays non établi dans la noticeOrganisme public
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Commissariat à l'Énergie Atomique et aux Énergies Alternatives pays non établi dans la noticeOrganisme public
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Université Paris-Saclay Département Médicaments et Technologies pour la Santé pays non établi dans la noticeUniversité ou école supérieure
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CEA Paris-Saclay pays non établi dans la noticeOrganisme public
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Infectious Disease Models and Innovative Therapies pays non établi dans la noticeStructure de recherche
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Institut National de Recherche pour l'Agriculture pays non établi dans la noticeOrganisme public
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NG Biotech (France) pays non établi dans la noticeEntreprise
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Médicaments et Technologies pour la Santé pays non établi dans la noticeStructure de recherche
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Bicêtre Hospital Department of Bacteriology-Hygiene pays non établi dans la noticeÉtablissement de santé
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CNR de la Résistance aux Antibiotiques pays non établi dans la noticeStructure de recherche
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Faculty of Medicine UMR1184 ‘Infectious Disease Models and Innovative Therapies (IDMIT)’ INSERM pays non établi dans la noticeUniversité ou école supérieure
Department of Pathobiology and Epidemiology — University of Sarajevo, Inserm et Commissariat à l'Énergie Atomique et aux Énergies Alternatives, avec 9 autres affiliations.
Une affiliation ne permet pas de déduire la nationalité d’un auteur.