Multiplex reverse transcriptase-polymerase chain reaction applied to virusdetection in globe artichoke

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.