Analytical Evaluation of KarbaDia, a Multiplex Immunochromatographic Assay for Rapid Detection of Major Carbapenemase Families in Gram-Negative Bacteria

Background: Rapid detection of carbapenemase-producing Gram-negative bacteria is critical for timely antimicrobial stewardship and infection control interventions. This study evaluated the analytical diagnostic performance of KarbaDia as a new multiplex lateral flow immunochromatographic assay for rapid detection of KPC, NDM, VIM, IMP, and OXA-۴۸-like carbapenemases. Materials & Methods: An analytical diagnostic accuracy evaluation study was conducted using ۷۸ non-duplicate well-characterized Gram-negative bacterial isolates obtained from publicly available diversity panels. The collection included carbapenemase-producing and carbapenemase-negative isolates of Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii. Whole genome sequencing (WGS) served as the reference standard. KarbaDia (GaDia SA, Switzerland) was evaluated in parallel with the Carbapenemase Detection Kit (Colloidal Gold; Macro & Micro-Test). Sensitivity, specificity, and ۹۵% confidence intervals (CI) were calculated for each carbapenemase family and for overall assay performance. Findings: KarbaDia demonstrated ۱۰۰% sensitivity for KPC (۶ of ۶), NDM (۱۳ of ۱۳), VIM (۳ of ۳), and IMP (۲ of ۲) and ۹۰.۰% for OXA-۴۸-like enzymes (۹ of ۱۰). The overall sensitivity was ۹۷.۱% (۹۵% CI: ۸۵.۱–۹۹.۹), while the specificity reached ۱۰۰% (۹۵% CI: ۹۹.۰–۱۰۰). No false-positive results were observed. The comparator assay demonstrated similar overall sensitivity (۹۷%) but produced four invalid results (۴.۸%), mainly associated with viscous K. pneumoniae colonies and incomplete chromatographic migration. Conclusion: KarbaDia demonstrated high analytical accuracy for rapid and multiplex detection of major carbapenemase families. The assay combines rapid turnaround time, multiplex capability, and ease of visual interpretation without dedicated instrumentation. These characteristics support its potential applicability in routine clinical microbiology laboratories; however, prospective multicenter clinical validation studies remain necessary before routine implementation.

کلیدواژه ها:

Carbapenemases ، Drug resistance ، Immunochromatography ، Klebsiella pneumoniae ، Pseudomonas aeruginosa ، Acinetobacter baumannii