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Oral: Isothermal Pathogen Detection


Development of a rapid AmplifyRP Acceler8 test for grapevine red blotch-associated virus in grapevines
R. LI (1), M. Fuchs (2), K. Perry (3), T. Mekuria (4), S. Zhang (1) (1) Agdia Inc., U.S.A.; (2) Cornell University, U.S.A.; (3) Cornell University, U.S.A.; (4) Vintage Nurseries, U.S.A.; (5) Agdia Inc., U.S.A.

Grapevine red blotch-associated virus (GRBaV) is of unprecedented concern to the grape industry since its discovery in 2011. A rapid, reliable, sensitive, and user-friendly test is needed to quickly identify GRBaV-infected grapevines and facilitate their timely removal from vineyards. An AmplifyRP Acceler8 detection kit was developed for GRBaV for use both in laboratories and vineyards. The test is performed in a disposable detection chamber for 20 minutes using crude grapevine extracts. The test specifically detects up to 11 copies of GRBaV genomic DNA in a matrix of healthy crude extract. The test has no cross reactivity to host plant tissues and grapevine-infecting pathogens including arabis mosaic virus (ArMV), grapevine fanleaf virus (GFLV), grapevine leafroll-associated virus 1 (GLRaV-1), grapevine leafroll-associated virus 2 (GLRaV-2), grapevine leafroll-associated virus 3 (GLRaV-3), grapevine leafroll-associated virus 4 strain 5 (GLRaV-4 strain 5), grapevine fleck virus (GFkV), tomato ringspot virus (ToRSV), tobacco ringspot virus (TRSV), Xylella fastidiosa, and Botrytis cinerea. The pellets contain all the necessary reaction components and are stable for a long period of storage even at 37°C. The test has been validated using both viral DNA and crude plant extracts as templates.