Abstract：
:Pollen transmitted viruses require accurate detection and identification to minimize the risk of spread through the global import and export of pollen. Therefore in this study we developed RT-qPCR assays for the detection of Cherry leaf roll virus (CLRV), Prune dwarf virus (PDV), Prunus necrotic ringspot virus (PNRSV), and Cherry virus A (CVA), four viruses that infect pollen of Prunus species. Assays were designed against alignments of extant sequences, optimized, and specificity was tested against known positive, negative, and non-target controls. An examination of assay sensitivity showed that detection of virus at concentrations as low as 101 copies was possible, although 102 copies was more consistent. Furthermore, comparison against extant assays showed that in both pollen and plant samples, the newly developed RT-qPCR assays were more sensitive and could detect a greater range of isolates than extant endpoint RT-PCR and ELISA assays. Use of updated assays will improve biosecurity protocols as well as the study of viruses infecting pollen.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Beaver-Kanuya E,Harper SJdoi
10.1016/j.jviromet.2019.113673subject
Has Abstractpub_date
2019-09-01 00:00:00pages
113673eissn
0166-0934issn
1879-0984pii
S0166-0934(19)30185-5journal_volume
271pub_type
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