Purification of potato virus A and its detection in potato by enzyme-linked immunosorbent assay (ELISA)

Abstract

Potato virus A (PVA) was purified fromNicandra physaloides by a simple method that omitted organic solvent clarification and consisted of differential centrifugation followed by equilibrium centrifugation in CsCl. An antiserum was produced that specifically detected PVA in potato leaf sap using either the SDS-agar test or enzyme-linked immunosorbent assay (ELISA). No heterlogous reaction of the antiserum with potato virus Y was detected. Purified PVA was highly infectious; it had an A 258/280 nm absorbance ratio of 1.28. The particles had a normal intact appearance in the electron microscope. Detection of PVA in potato sprouts and foliage by ELISA was compared with the local lesion assay onPhysalis angulata L. plants. ELISA was superior over an indicator plant test when sprout tissue was used. PVA antiserum reacted similarly with mild and crinkle isolates.

Resumen

El virus A de la papa (PVA) fue purificado deNicandra physaloides por un método simple donde se omitió la clarificación con solvente orgánico y consistió de centrifugación diferencial seguida por centrifugación hasta equilibrio en Cs Cl. Se preparó un antisuero que detectó específicamente PVA en hojas de papa usando la prueba de difusión an agar-SDS ó la técnica serológica con conjugado enzimático (ELISA). No se detectó reacción heteróloga del antisuero con el virus y de la papa. PVA purificado fue altamente infeccioso; la relación A 260/280 tuvo una absorbancia de 1.28. Las partículas aparecían normales e intactas en el microscopio electrónico. Detección de PVA en brotes y follaje de papa por medio de ELISA fue comparada con la producción de lesiones locales enPhysalis angulata L. ELISA fue superior que la planta indicadora cuando se probó en brotes. El antisuero de PVA reaccionó similarmente con aislamientos suaves y con aquellos que producen encrespamiento.

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Singh, R.P., McDonald, J.G. Purification of potato virus A and its detection in potato by enzyme-linked immunosorbent assay (ELISA). American Potato Journal 58, 181–189 (1981). https://doi.org/10.1007/BF02854418

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Key Words

  • Potato virus Y
  • serology