POSTERS: Pathogen detection, quantification and diagnosis
Development of AmplifyRP assays for three important grapevine viruses through recombinase polymerase amplification
Rugang Li - Agdia, Inc.. Shulu Zhang- Agdia, Inc., Deborah Groth-Helms- Agdia, Inc., Bryant Davenport- Agdia, Inc.
Focusing on a higher level of sensitivity, the specificity, and the simplicity of detection application, Agdia has developed three virus-specific AmplifyRP® XRT assays for three important grapevine viruses: Grapevine leafroll associated virus 3 (GLRaV-3), Grapevine pinot gris virus (GPGV), and Grapevine red blotch virus (GRBV). For each assay, all reaction reagents are lyophilized into a single pellet in a PCR tube and rehydrated in a buffer prior to adding testing sample. Crude extracts are prepared by grinding leaf petiole or leaf base in 1:10 extraction buffer ratio (wt/vol) in extraction bags and directly used for testing. Assays can be completed within 20 minutes in a portable reader, AmpliFire®. AmplifyRP® XRT for GLRaV-3 can detect the GLRaV-3 variant representatives from eight known groups (I, II, III, V, VI, VII, IX and X) while excluding other grapevine leafroll viruses. AmplifyRP® XRT for GPGV can detect 27 GPGV isolates collected from Italy, Canada, and USA without cross-reacting to other grapevine viruses. AmplifyRP® for GRBV was previously developed as the Acceler8® format and recently the XRT format was generated. AmplifyRP® XRT has demonstrated to be as sensitive as RT-PCR or PCR. Both GLRaV-3 and GPGV can be detected from crude infected plant extracts diluted to 1:104 while GRBV can be detected in crude extracts diluted to 1:106. These new assays provide convenient and fast tools for growers to monitor key viruses in their vineyards.