Use of polymerase chain reaction and sandwich-hybridization for detection of artichoke mottled crinkle tombusvirus in artichoke

Two non-radioactive methods were applied for the detection of artichoke mottled crinkle tombusvirus (AMCV) in artichoke plant sap. A sandwich hybridization method was developed using a biotinylated transcript as capture and a digoxigenin-labelled riboprobe. The hybrid was collected onto Nylon membranes coated with streptavidin using a slot-blot procedure. The second method was the reverse transcriptional polymerase chain reaction (RT-PCR) carried out with three different primers obtained from widely, separated sequences of AMCV genome. In artichoke plant sap, RT-PCR was 20-fold more sensitive than sandwich-hybridization and 2.5-fold less sensitive than dor-blot hybridization labelled with radioactive reporter groups.