F. Grieco et D. Gallitelli, Multiplex reverse transcriptase-polymerase chain reaction applied to virusdetection in globe artichoke, J PHYTOPATH, 147(3), 1999, pp. 183-185
Citations number
11
Categorie Soggetti
Plant Sciences
Journal title
JOURNAL OF PHYTOPATHOLOGY-PHYTOPATHOLOGISCHE ZEITSCHRIFT
Six specific primers were used for reverse transcription-amplification of p
arts of the genomic RNAs of artichoke mottled crinkle tombusvirus (AMCV), a
rtichoke Italian latent nepovirus (AILV) and artichoke latent potyvirus (AL
V) in artichoke plant sap. Each primer pair was highly specific for each vi
ral RNA template and in multiplex reverse transcription-amplification react
ions run independently, yielding products of the expected virus-specific si
ze. The optimal artichoke plant sap dilution was about 10(3)-fold in 10 mM
DTT and the optimal MgCl2 concentration was 2.25 mM. The detection limits f
or simultaneous amplification were of about 400 fg for AMCV and ALV and of
about 4 pg for AILV. The method proved to be useful for simultaneous detect
ion in cases of viral mixed infections which are likely to occur in nature.