Real-time NASBA for sensitive detection of Strawberry vein banding virus

An assay for the detection of Strawberry vein banding virus (SVBV) in Fragaria spp. based on nucleic acid sequence based amplification (NASBA) and real-time detection using molecular beacons (real-time NASBA) is described. This assay was compared both with biological indexing, the current method for certification of SVBV, and a newly optimised PCR-based detection method. Performance of the assay was tested on three SVBV isolates in Fragaria indicator plants and 317 field strawberries from five European countries. The assay was shown to be SVBV-specific, testing negative for other common aphid-borne strawberry viruses. The virus was detected in purified total RNA preparations diluted a millionfold, which is an amount equivalent to 1ng of fresh material. The real-time NASBA method developed here offers the potential of a fast, sensitive and reliable approach for the routine diagnosis of strawberry stock material