Abstract

Mixed viral infections are common in globe artichoke (Cynara cardunculus var. scolymus). Despite lack ofevident symptoms, production losses are relevant. A rapid and accurate detection is advisable for selection and marketingof plant propagation material.Objectives: Our aim was the evaluation of a suspension microbeads array system for simultaneous detection ofArtichoke latent potyvirus (ArLV), Artichoke Italian latent nepovirus (AILV), Artichoke mottled crinkle tombusvirus (AMCV),Tomato spotted wilt tospovirus (TSWV), Turnip mosaic potyvirus (TuMV), Cucumber mosaic cucumovirus (CMV), Potatovirus X (PVX), Tobacco mosaic tobamovirus (TMV) and Pelargonium zonate spot anulavirus (PZSV).Methods: For PCR, oligonucleotides were designed from conserved regions in the viral genomes, using an Illumina´sAssay Design Tool. The actin gene of C. cardunculus was used as control. The microbeads-based procedure started witha first PCR target amplification. A further target-specific primer extension (TSPE), holding a unique 5' motif, was thenperformed. During this step biotinylated dCTPs were incorporated into the extension products. These were then linked tothe holographic microbeads bearing an oligo complementary to the 5' motif. Probe labelling was performed withstreptavidin-Alexa-647, for final scan using an Illumina BeadXpress Reader.Conclusions: Multiplex fluorescent microbeads assays allowed the contemporary detection of several viruses in a singlesample. This highly flexible microarray approach used a suspension of barcoded microbeads bearing specific sequencemotifs. It favoured the tailoring of user defined applications. The method is also suitable for identification of SNPs in viralstrains, within a unique host phenotype.

Autore Pugliese

• Finetti Sialer Mariella Matilde , Minafra Angelantonio

Tutti gli autori

• Finetti-Sialer M.M.; De Miccolis Angelini R.M.; Minafra A.

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Anno di pubblicazione

2013

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