| Abstract [eng] |
Bacterial resistance to antibiotics is a growing threat to human health globally. The widespread use of antimicrobials in the community, healthcare settings, and animal farming facilitate the rapid emergence and dissemination of antibiotic resistance mechanisms in bacteria. Currently, approximately 60% of all antibiotics administered in healthcare settings belong to the β-lactams. Therefore, β-lactamases – bacterial enzymes hydrolyzing this class of antibiotics – can serve as diagnostic targets. Monoclonal antibodies (MAbs) specific to β-lactamases can be applied for detection of these resistance factors in bacterial isolates and serve as a promising diagnostic tool for epidemiological studies and profiling of resistance mechanisms. This study aimed to generate MAbs against bacterial AmpC and metallo β-lactamases, for potential diagnostic applications. It describes MAbs raised against recombinant ACT-14, NDM-1, CMY-34 and PDC-195 β-lactamases, their comprehensive characterization and application in various immunoassays. Moreover, this study details the development and characterization of novel, broadly-reactive MAbs raised against a conserved 17 amino acid-long sequence of AmpC β-lactamases. To facilitate the production of immunogen, a novel approach utilizing bacteriophage-derived nanotubes displaying selected universal sequence of AmpC β-lactamases was employed. The observed reactivity of the MAbs with natural β-lactamases confirmed their diagnostic potential. |
