AUTHOR=Ivagnes Vittorio , De Maio Flavio , Baccani Ilaria , Antonelli Alberto , Menchinelli Giulia , Rosato Roberto , Cafaro Giordana , Santarelli Giulia , Falletta Federico , D’Inzeo Tiziana , Sanguinetti Maurizio , Spanu Teresa , De Angelis Giulia , Rossolini Gian Maria , Posteraro Brunella 

TITLE=Detection of β-lactam resistance genes in Gram-negative bacteria from positive blood cultures using a microchip-based molecular assay

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=Volume 15 - 2025

YEAR=2025

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2025.1597700

DOI=10.3389/fcimb.2025.1597700

ISSN=2235-2988

ABSTRACT=BackgroundAccurate detection of β-lactam resistance genes in bloodstream infections is critical for guiding antimicrobial therapy. This study evaluates the Alifax Gram-negative resistance (GNR) microchip assay for detecting β-lactam resistance genes directly from positive blood cultures (PBCs) for Gram-negative (GN) bacteria, including Enterobacterales, Pseudomonas aeruginosa, and Acinetobacter baumannii.MethodsSimulated (n=146) and clinical (n=106) GN-PBC samples were tested for blaKPC, blaVIM, blaNDM, blaIMP, blaOXA-23-like, blaOXA-48-like, blaSHV-ESBL, blaCTX-M-1/9 group, and blaCMY-2-like genes using the GNR microchip assay. Whole-genome sequencing (WGS) served as the reference assay for simulated samples and, selectively, for clinical samples. The bioMérieux BioFire Blood Culture Identification 2 (BCID2) panel assay was used as a comparator for clinical samples.ResultsThe GNR microchip assay correctly identified 203 (99.5%) of 204 β-lactam resistance genes in simulated samples. One sample tested false negative for a blaSHV-ESBL gene but true positive for a blaKPC gene. In clinical samples, GNR results were concordant with BCID2 for 113 (100%) of 113 genes included in both assays. Additionally, the GNR assay detected blaCMY-2-like (n=6), blaOXA-23-like (n=5), and blaSHV-ESBL (n=2), which are not targeted by BCID2, all confirmed by WGS. In two β-lactam-resistant P. aeruginosa samples but negative by the GNR assay, WGS confirmed the absence of acquired β-lactam resistance genes, suggesting alternative resistance mechanisms.ConclusionThe GNR microchip assay demonstrated high concordance and broader β-lactam resistance gene coverage compared to BCID2, supporting its potential role in routine diagnostics. Further validation in larger, prospective studies is warranted.