Simple and portable magnetic immunoassay for rapid detection and sensitive quantification of plant viruses. 2015

Stefanie Rettcher, and Felicitas Jungk, and Christoph Kühn, and Hans-Joachim Krause, and Greta Nölke, and Ulrich Commandeur, and Rainer Fischer, and Stefan Schillberg, and Florian Schröper
Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Aachen, Germany.

Plant pathogens cause major economic losses in the agricultural industry because late detection delays the implementation of measures that can prevent their dissemination. Sensitive and robust procedures for the rapid detection of plant pathogens are therefore required to reduce yield losses and the use of expensive, environmentally damaging chemicals. Here we describe a simple and portable system for the rapid detection of viral pathogens in infected plants based on immunofiltration, subsequent magnetic detection, and the quantification of magnetically labeled virus particles. Grapevine fanleaf virus (GFLV) was chosen as a model pathogen. Monoclonal antibodies recognizing the GFLV capsid protein were immobilized onto immunofiltration columns, and the same antibodies were linked to magnetic nanoparticles. GFLV was quantified by immunofiltration with magnetic labeling in a double-antibody sandwich configuration. A magnetic frequency mixing technique, in which a two-frequency magnetic excitation field was used to induce a sum frequency signal in the resonant detection coil, corresponding to the virus concentration within the immunofiltration column, was used for high-sensitivity quantification. We were able to measure GFLV concentrations in the range of 6 ng/ml to 20 μg/ml in less than 30 min. The magnetic immunoassay could also be adapted to detect other plant viruses, including Potato virus X and Tobacco mosaic virus, with detection limits of 2 to 60 ng/ml.

UI MeSH Term Description Entries
D010935 Plant Diseases Diseases of plants. Disease, Plant,Diseases, Plant,Plant Disease
D013997 Time Factors Elements of limited time intervals, contributing to particular results or situations. Time Series,Factor, Time,Time Factor
D014027 Tobacco Mosaic Virus The type species of TOBAMOVIRUS which causes mosaic disease of NICOTIANA. Transmission occurs by mechanical inoculation. Mosaic Virus, Tobacco,Mosaic Viruses, Tobacco,Tobacco Mosaic Viruses,Virus, Tobacco Mosaic,Viruses, Tobacco Mosaic
D053758 Nanoparticles Nanometer-sized particles that are nanoscale in three dimensions. They include nanocrystaline materials; NANOCAPSULES; METAL NANOPARTICLES; DENDRIMERS, and QUANTUM DOTS. The uses of nanoparticles include DRUG DELIVERY SYSTEMS and cancer targeting and imaging. Nanocrystalline Materials,Nanocrystals,Material, Nanocrystalline,Materials, Nanocrystalline,Nanocrystal,Nanocrystalline Material,Nanoparticle
D017860 Nepovirus A genus of polyhedral plant viruses of the family COMOVIRIDAE causing ringspots and spotting on leaves or sometimes symptomless infection. Transmission occurs by seeds, soil nematodes, or experimentally by mechanical inoculation. Tobacco ringspot virus is the type species. Tobacco ringspot virus,Nepoviruses,Tobacco ringspot viruses
D017863 Potexvirus A genus of plant viruses in the family FLEXIVIRIDAE, that cause mosaic and ringspot symptoms. Transmission occurs mechanically. Potato virus X is the type species. Potato virus X,Potexviruses
D018189 Immunomagnetic Separation A cell-separation technique where magnetizable microspheres or beads are first coated with monoclonal antibody, allowed to search and bind to target cells, and are then selectively removed when passed through a magnetic field. Among other applications, the technique is commonly used to remove tumor cells from the marrow (BONE MARROW PURGING) of patients who are to undergo autologous bone marrow transplantation. Immunomagnetic Bead Technique,Immunomagnetic Purging,Immunomagnetic Cell Separation,Bead Technique, Immunomagnetic,Bead Techniques, Immunomagnetic,Cell Separation, Immunomagnetic,Cell Separations, Immunomagnetic,Immunomagnetic Bead Techniques,Immunomagnetic Cell Separations,Immunomagnetic Purgings,Immunomagnetic Separations,Purging, Immunomagnetic,Purgings, Immunomagnetic,Separation, Immunomagnetic,Separation, Immunomagnetic Cell,Separations, Immunomagnetic,Separations, Immunomagnetic Cell
D019562 Viral Load The quantity of measurable virus in a body fluid. Change in viral load, measured in plasma, is sometimes used as a SURROGATE MARKER in disease progression. Viral Burden,Virus Titer,Burden, Viral,Load, Viral,Titer, Virus

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