SIMULTANEOUS DETECTION OF CUCUMBER MOSAIC-VIRUS, TOMATO MOSAIC-VIRUS AND POTATO-VIRUS-Y BY FLOW-CYTOMETRY

The simultaneous detection is described of cucumber mosaic virus (CMV) , potato virus Y (PVY) and tomato mosaic virus (ToMV) by flow cytometr y. Extracts from leaves of healthy and CMV or PW infected plants were incubated with latex particles, each with a diameter of 3 mu m. Extrac ts from ToMV infected or uninfected plants, however, were incubated wi th particles, each with a diameter of 6 mu m. Beads were washed and in cubated in succession with primary and secondary antibodies, the latte r labeled with phycoerythrin (PE) or fluorescein (FITC). CMV and PVY w ere distinguished on the basis of the fluorescence emitted by FITC and PE; ToMV was distinguished from CMV and PVY on the basis of the diffe rent diameter (6 mu m) of the particles on which it was adsorbed. The three viruses were detected also by another approach. Latex particles with a diameter of 3, 6 and 10 mu m were separately sensitized with an tibodies specific for CMV, PVY and ToMV. An equal number of sensitized particles was mixed and incubated with the plant extracts containing the three viruses and then with anti-CMV, anti-PVY and anti-ToMV antib odies labeled with FITC. The study describes also a virus purification method based on the use of antibody coated latex particles. The metho d is simple technically and applicable to the purification of large as well as minute amounts of different viruses (CMV, PVY and ToMV). (C) 1997 Elsevier Science B.V.