The planned project will be dedicated to a pathogen-specific detection and will enable a POC application for veterinary issues. The basic idea to detect the immunogen directly by labeled antibodies will be maintained in the present project. The knowledge gained from other projects will be used for the direct detection of pathogenic bacteria and will also be expanded in the multiplex procedure for the detection of antibiotic resistance. The ubiquitous bacterium Escherichia coli, which is to be detected using antibodies already developed, will initially serve as a model.
IIn a two-step procedure, the pathogen is first detected and then digested to access intracellular resistance-mediating proteins. For this purpose, magnetic beads with covalently bound pathogen-specific antibodies are fixed on a magnetizable surface after incubation with the sample and detected by fluorescently labeled pathogen-specific antibodies. By application of lysing detergents or enzymes, the intracellular proteins are released and - if present - captured by specific camelid antibodies against ß-lactamase predoped onto the surface. Finally, detection is performed by fluorescence-labeled camelid full-length antibodies.