Lateral flow immunoassay for simultaneous detection of C. difficile, MRSA, and K. pneumoniae

Mainly performed within a rapid diagnostic tests company, a lateral flow (LF) system using gold nanoparticles (AuNPs) as transducers is presented able to detect three bacteria of interest, of relevance for antimicrobial resistance (AMR): Clostridioides difficile, methicillin-resistant Staphylococcus aureus (MRSA), and Klebsiella pneumoniae, with a limit of detection of 25 ng/mL of glutamate dehydrogenase (GDH) for C. difficile, 36 ng/mL of penicillin-binding protein 2a (PBP2a) for MRSA, and 4 × 10⁶ CFU/mL for K. pneumoniae. The system showed good results with bacteria culture samples, is user-friendly, and suitable for rapid testing, as the results are obtained within 15 min.