[Investigation of electrophysical properties of Listeria monocytogenes cells during the interaction with monoclonal antibodies]

An electrooptical approach was used in studies of Listeria monocytogenes-antibody binding. An electrooptical analyzer, which has been developed at the State Research Center for Applied Microbiology (Obolensk, Russia), was used as a basic instrument for electrooptical measurements. The analyzer consists of the following modules: a sample preparation module, a mixer, an AC field generator, an EO-flow cell, a microcontroller for transfer of liquid, a thermal system, an operator interface, and an image processor. The sample preparation module includes a unit for an automatic filter changing device and a hydraulic system. Since the AC electrokinetic effects depend on the dielectric properties of bioparticles, their composition, morphology, phenotype, the medium, and the frequency of applied electrical field, the electroorientational spectra were used for discrimination of different types of bacteria, a given type being "controlled" (and identified) by the selective choice of binding agents (antibodies). The measurements were performed using a discrete set of frequencies of the orienting electric field (10, 100, 250, and 500 kHz). During biospecific interactions, an antibody is bound to the microorganism, causing a change in the dielectric properties of the microorganism-antibody complex, and the electrooptic signal reaches its maximum at 100-200 kHz. It was shown that the biospecific interactions of Listeria monocytogenes cells with anti-Listeria antibody in the presence of E. coli K-12, and A. brasilense sp7 significantly change the electrooptical signals. Thus, the determination of the presence of particular bacteria within a mixed sample may be achieved by selection and matching of antibodies specific to individual bacterium types and by comparing the spectra of bacterium in the presence and in the absence of specific binding agent (antibody).