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Potato leafroll virus purification and antiserum preparation for enzyme-linked immunosorbent assays

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Abstract

Potato leafroll virus (PLRV) was purified from potato foliage and stems with an average yield of 0.14 mg of PLRV/kg of potato. Modifications of an existing purification procedure are reported. Five low dosage (38-118 μg of PLRV) intravenous injections were used to produce a PLRV antiserum for use in enzyme-linked immunosorbent assays (ELISA) from tubers. PLRV was readily detected in ELISA testing of potato tubers and leaves and inPhysalis floridana Rybd. Non-specific reactions were low with all tissues. In parallel tests, a Canadian antiserum produced higher nonspecific reactions with tuber and leaf tissue. The results indicated that the use of low dosage-intravenous injections might be necessary methodology for producing PLRV antiserum for use in ELISA diagnostic tests with tuber tissue where high non-specific reactions have been reported.

Resumen

El virus del enrollamiento de las hojas de papa (PLRV) fue purificado de follaje y tallos de papa con un rendimiento promedio de 0.14 mg. de PLRV/Kg. de papa. Se reportan modificaciones de un procedimiento de purificación. Cinco inyecciones intravenosas con dosis bajas (38-118 μg de PLRV) se usaron para producir un antisuero para PLRV para ser usado en pruebas de immuno-absorción con conjugado enzimático (ELISA) en tubérculos. PLRV fue detectado fácilmente por ELISA en tubérculos y hojas de papa, así como dePhysalis floridana Rybd. Reacciones inespecíficas fueron bajas con todos los tejidos. En pruebas paralelas, un antisuero canadiense produjo reacciones inespecíficas más altas con tejido de tubérculos y hojas. Los resultados indicaron que el uso de inyecciones intravenosas con dosis bajas podria ser la metodologia necesaria para producir antisueros a PLRV para uso en pruebas con ELISA con tejido de tubérculos donde se han reportado reacciones inespecificas altas.

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Authors and Affiliations

  1. Department of Entomology, Oregon State University, 97331, Corvallis, OR

    R. G. Clarke (Assistant Professor)

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  1. R. G. Clarke
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Oregon Agricultural Experiment Station Technical Paper No. 5652.

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Clarke, R.G. Potato leafroll virus purification and antiserum preparation for enzyme-linked immunosorbent assays. American Potato Journal 58, 291–298 (1981). https://doi.org/10.1007/BF02854095

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  • DOI: https://doi.org/10.1007/BF02854095

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