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Anwendung der Pofymerase‐Kettenreaktion und der Sandwich‐Hybridisierung bei der Ermittlung des artichoke mottled crinkle Tombusvirus in Artischocken
Author(s) -
Barbarossa L.,
Grieco F.,
Iosco P.,
Gallitelli D.
Publication year - 1994
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/j.1439-0434.1994.tb04809.x
Subject(s) - riboprobe , biology , jerusalem artichoke , polymerase chain reaction , digoxigenin , microbiology and biotechnology , dot blot , biotinylation , virology , gene , in situ hybridization , messenger rna , genetics , biochemistry
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.