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Viruses of Ullucus tuberosus: The opportunities and implications of using next generation sequencing in support of statutory diagnostics

Adrian Fox: Fera Science Ltd

<div>Ulluco (<em>Ullucus tuberosus</em>) is a tuber forming crop routinely grown alongside potatoes in the South American Andes, which has become a novel crop in both highland and temperate maritime climates. Eight viruses have been previously reported infecting ulluco, including a strain of <em>Andean potato latent virus </em>(APLV). Ulluco is not currently listed as regulated in EU trade. Following a breach of UK plant health regulations, 400 ulluco plants from a small holding in South-West England were tested for the presence of quarantine viruses using ELISA and real-time PCR. Following positive ELISA results for APLV and multiple other viruses, virus identification was confirmed by Next Generation Sequencing (NGS) using a ribosomal RNA (rRNA) depleted total RNA approach on an Illumina MiSeq platform. Analysis of viral contigs indicated the presence of several novel viruses closely related to, but not consistent with, the viruses indicated by ELISA or those previously described from ulluco. A second outbreak site linked to internet trade in ulluco tubers was investigated using the same approach as described and a similar but distinct suite of viruses was detected. Using NGS in a diagnostic confirmation role offers the ability to identify multiple viruses simultaneously, whilst alleviating the concerns over novel findings without supporting diagnostic context. The limitations of list-based regulatory systems in the era of internet trade will be highlighted.</div>

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