Premium
Use of fluorogenic Scorpions for fast and sensitive detection of plant viruses *
Author(s) -
Sialer M.M. Finetti,
Ciancio A.,
Gallitelli D.
Publication year - 2000
Publication title -
eppo bulletin
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.327
H-Index - 36
eISSN - 1365-2338
pISSN - 0250-8052
DOI - 10.1111/j.1365-2338.2000.tb00925.x
Subject(s) - amplicon , biology , primer (cosmetics) , ethidium bromide , fluorescence , microbiology and biotechnology , nucleic acid , virology , chromatography , dna , chemistry , polymerase chain reaction , biochemistry , gene , physics , organic chemistry , quantum mechanics
Very recently, a novel nucleic acid detection technology based on a primer with a quenched fluorophore and a probe complementary to an internal portion of the amplicon has been developed. The primer probe, named ‘Scorpion’, fluoresces only upon hybridization with its target, i.e. if the specific extension product of the primer is synthesized during the reaction. We have used Scorpion primers in RT‐PCR assays to detect Tomato spotted wilt tospovirus and Plum pox potyvirus in naturally infected plants. Amplified products of the expected size were detected either by real‐time measurements or by visual estimation of fluorescence emission from tubes placed over an ultraviolet light source at the end of the Scorpion‐RT‐PCR. In serially diluted plant sap, the visual method allowed detection of 7.5 pg of infected plant tissue and proved at least as sensitive as visual estimation of ethidium bromide‐stained amplicons separated by gel electrophoresis.