title: Development of a portable, high throughput biosensor system for rapid plant virus detection.
abstract: Biosensors based on living cells are characterized by high sensitivity, selectivity and rapid response times. A novel portable cell biosensor system for the detection of plant viruses, based on immobilized 'Vero' cells carrying on their membrane virus specific antibodies was developed and was designated as High Throughput Bioelectric Recognition Assay (BERA-HTP). BERA-HTP was tested for the detection of purified Potato virus Y (PVY), Cucumber mosaic virus (CMV) and Tobacco rattle virus (TRV) and of CMV and TRV in single, as well as in mixed infections in two different plant host species. The sensor was based on live, mammalian cells, the membrane of which has been artificially saturated with antibodies specific to different plant viruses. The attachment of PVY, CMV or TRV viral particles to the homologous electroinserted antibodies caused a virus-specific change of the cell membrane electric potential that was not observed with virus-free samples or with heterologous viruses. Fluorescence microscopy observations showed that attachment of virus particles to the cell membrane bearing the homologous antibody, was associated with a decrease of [Ca(2+)]cyt. The perspective for the development of BERA-HTP as a portable, reliable and rapid (duration of assay for 96 samples: ∼70 min) detection method of plant viruses in the field is discussed.
authors.affiliation: Laboratory of Plant Physiology, Faculty of Biotechnology, Agricultural University of Athens, Iera Odos 75, 118 55 Athens, Greece.
authors.forename: Antonios
authors.initials: A
authors.lastname: Perdikaris
authors.forename: Nikon
authors.initials: N
authors.lastname: Vassilakos
authors.forename: Iakovos
authors.initials: I
authors.lastname: Yiakoumettis
authors.forename: Oxana
authors.initials: O
authors.lastname: Kektsidou
authors.forename: Spiridon
authors.initials: S
authors.lastname: Kintzios
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation; D002522:Chlorocebus aethiops; D057166:High-Throughput Screening Assays / Q000295:instrumentation; D054884:Host-Pathogen Interactions; D008856:Microscopy, Fluorescence; D010942:Plant Viruses / Q000302:isolation & purification*; D014026:Nicotiana / Q000821:virology; D014709:Vero Cells