{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T15:39:33Z","timestamp":1792510773162,"version":"4.5.0"},"reference-count":24,"publisher":"American Society for Microbiology","issue":"10","license":[{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"National Reference Center for Emerging Antibiotic Resistance (NARA), Fribourg, Switzerland"},{"DOI":"10.13039\/501100005869","name":"Universit\u00e9 de Fribourg","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100005869","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2026,10,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      A chromogenic biochemical assay (ESBL-Carba Combo PN test) was developed for the simultaneous detection of extended-spectrum \u03b2-lactamase (ESBL) and carbapenemase activity in Enterobacterales, as well as for discrimination of clinically relevant carbapenemase classes. The assay is based on a novel chromogenic substrate, B-Lac, and a low-cost Tris-based lysis buffer, providing a unified workflow for the detection of ESBLs and carbapenemases. The test was evaluated using cultures of well-characterized clinical isolates with resistance mechanisms previously defined by whole-genome sequencing. Results were obtained after a mean preparation time of approximately 3 min per isolate and were read after 20 min and 1 h of incubation. For ESBL detection (\n                      <jats:italic toggle=\"yes\">n<\/jats:italic>\n                      = 67), sensitivity was 93.8% and 97.8% after 20 min and 1 h of incubation, respectively, with a specificity of 100%. Carbapenemase activity (\n                      <jats:italic toggle=\"yes\">n<\/jats:italic>\n                      = 92) was detected with 100% sensitivity and specificity at both time points. Using the combined assay (\n                      <jats:italic toggle=\"yes\">n<\/jats:italic>\n                      = 85), ESBL and\/or carbapenemase activity was detected with 100% sensitivity and specificity. For Ambler classes A and B, 100% sensitivity and specificity were reached after 20 min of incubation. Regarding class D carbapenemases, 91.3% and 95.5% sensitivity after 20 min and 1 h of incubation, respectively, were observed, with 100% specificity at both times. The ESBL-Carba Combo PN test provides a simple, low-cost (&lt;1 USD\/sample), and functional method for the simultaneous detection of the major \u03b2-lactam resistance mechanisms in Enterobacterales and for carbapenemase class discrimination within one workflow, potentially supporting earlier optimization of antimicrobial therapy and antimicrobial stewardship in routine clinical microbiology laboratories.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>Rapid identification of extended-spectrum \u03b2-lactamase (ESBL)- and carbapenemase-producing Enterobacterales (CPE) is essential to optimize antimicrobial therapy and implement timely infection control measures. Most currently available rapid phenotypic assays detect these resistance mechanisms separately, increasing hands-on time, workload, reagent consumption, and cost, particularly in isolates co-producing multiple \u03b2-lactamases. In addition, discrimination between carbapenemase classes, particularly class A and class D carbapenemases, remains challenging for most biochemical assays. This study describes a simple chromogenic assay that allows the simultaneous functional detection of both enzyme types using a single substrate and a low-cost lysis buffer. By incorporating relebactam as a selective class A inhibitor, the assay improves phenotypic discrimination between class A and class D carbapenemases while preserving reliable ESBL detection. The rapid ESBL-Carba Combo PN test provides 1 h of phenotypic information on the most clinically relevant \u03b2-lactam resistance mechanisms and represents a useful tool to guide early antibiotic treatment.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.01123-26","type":"journal-article","created":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T16:45:50Z","timestamp":1788367550000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Rapid ESBL-Carba Combo PN test for simultaneous detection of ESBL and carbapenemase activity in Enterobacterales"],"prefix":"10.1128","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8019-0905","authenticated-orcid":true,"given":"Soraya","family":"Herrera-Espejo","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/022fs9h90","id-type":"ROR","asserted-by":"publisher"}],"name":"Medical and Molecular Microbiology, Faculty of 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