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 dot-blot hybridization labelled with radioactive reporter groups.
USE OF THE POLYMERASE CHAIN-REACTION AND SANDWICH-HYBRIDIZATION FOR DETECTING ARTICHOKE MOTTLED CRINKLE TOMBUSVIRUS IN ARTICHOKE
GALLITELLI, Donato
1994-01-01
Abstract
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 dot-blot hybridization labelled with radioactive reporter groups.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
