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ISHS Acta Horticulturae 127: IV Conference on recent Advances in Vegetable Virus Research

ADVANCES IN VIRUS IDENTIFICATION USING IMMUNOSORBENT ELECTRON MICROSCOPY

Author:   D.-E. Lesemann
DOI:   10.17660/ActaHortic.1983.127.12
Abstract:
Some parameters of the technique of immunosorbent electron microscopy (ISEM) and the application of ISEM for a highly sensitive and specific detection and identification of virus particles is described and discussed. The highest sensitivity is achieved by long incubation times of several hours of antiserum coated grids (ACGs) with the virus suspensions. The sensitivity is at limiting virus concentrations not influenced by an increase of antibody concentration on ACGs. However, the specificity of ISEM depends on the density of antibody coating on ACG and on the incubation times with the virus suspension. Homologous reactions are clearly differentiated with antiserum diluted c. 1:1000, and short incubation times of c. 75 min. Weak heterologous reactions are clearly detected on grids with high antibody density and after long incubation times, e.g. 2 hours. A dense layer of antibodies is achieved by first coating grids with protein A which traps antibodies during subsequent incubation on little dilute antisera. The trapping reaction varies in wide ranges with different viruses. The reliability of the results with heterologous trapping is not yet enough evaluated.

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