Performance of Flow Cytometry-Based Rapid Assay in Detection of Carbapenemase-Producing Enterobacterales

• Published in: International journal of molecular sciences 2024-07, Vol.25 (14), p.7888
• Main Authors: Pérez-Viso, Blanca, Martins-Oliveira, Inês, Gomes, Rosário, Silva-Dias, Ana, Peixe, Luísa, Novais, Ângela, Pina-Vaz, Cidália, Cantón, Rafael
• Format: Article
• Language: English
• Subjects: Acids; Anti-Bacterial Agents - pharmacology; Antibacterial agents; Antibiotics; Antimicrobial agents; Bacterial infections; Bacterial Proteins - metabolism; Beta lactamases; beta-Lactamases - metabolism; Carbapenems - pharmacology; Enterobacteriaceae - drug effects; Enterobacteriaceae - enzymology; Enzymes; Ethylenediaminetetraacetic acid; Flow cytometry; Flow Cytometry - methods; Health aspects; Humans; Infection; Microbial Sensitivity Tests - methods; Mortality; Nosocomial infections; Oxalic acid; Public health; Sensitivity and Specificity; Spinoffs
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms25147888

Carbapenemase-producing Enterobacterales are increasingly being recognized in nosocomial infections. The performance of a flow cytometry-based rapid assay for their detection and differentiation was evaluated. This is a disruptive phenotypic technology, phenotypic and growth-independent, that searches for the lesions produced by drugs acting on cells after a short incubation time. Overall, 180 Gram-negative bacteria were studied, and results were compared with those obtained molecularly by PCR and phenotypically by 'KPC, MBL and OXA-48 Confirm Kit'. This phenotypic method was used as reference for comparison purposes. Susceptibility to carbapenems (imipenem, meropenem, and ertapenem) was determined by standard broth microdilution. Overall, 112 isolates (62.2%) were carbapenemase producers, 41 KPCs, 36 MβLs, and 31 OXA-48, and 4 strains were KPC + MβL co-producers. Sixty-eight isolates were carbapenemase-negative. The percentage of agreement, sensitivity, and specificity were calculated according to ISO 20776-2:2021. The FASTinov assay showed 97.7% agreement with the reference method for carbapenemase detection. Discrepant flow cytometry results were obtained in four isolates compared with both reference and PCR results. The sensitivity and specificity of this new technology were 95.3% and 98.5%, respectively, for KPCs, 97.6% and 99.3% for MβLs, and 96.9% and 98% for OXA-48 detection. In conclusion, we describe a rapid flow cytometry assay with high accuracy for carbapenemase detection and the differentiation of various carbapenemases, which should impact clinical microbiology laboratories and patient management.