Molecular diagnosis of grapevine fleck virus

A digoxigenin-labelled riboprobe was developed for the detection of grapevine fleck virus (GFkV) in infected tissues of grapevine leaves, roots and canes. The probe was GFkV-specific and was successfully used for virus identification both with dot spot assays, using alkali-treated crude sap, and tissue blot assays, using cross and longitudinal sections of leaf petioles. Primers designed for the amplification by reverse transcription-polymerase chain reaction of a viral genome fragment 243 nucleotides in size, gave also positive and repeatable results. These newly developed molecular-based detection tools extend the range of available procedures for the sensitive identification of GFkV in naturally infected hosts