Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 501731
Title Generic RT-PCR tests for detection and identification of tospoviruses
Author(s) Hassani-Mehraban, A.; Westenberg, M.; Verhoeven, J.T.J.; Vossenberg, B.T.L.H. van de; Kormelink, R.; Roenhorst, J.W.
Source Journal of Virological Methods 233 (2016). - ISSN 0166-0934 - p. 89 - 96.
DOI http://dx.doi.org/10.1016/j.jviromet.2016.03.015
Department(s) Laboratory of Virology
PPO/PRI Biointeractions and Plant Health
EPS
Publication type Refereed Article in a scientific journal
Publication year 2016
Keyword(s) Bunyaviridae - Clade-specific primers - Diagnostics - Nucleocapsid gene - Plant virus
Abstract

A set of tests for generic detection and identification of tospoviruses has been developed. Based on a multiple sequence alignment of the nucleocapsid gene and its 5' upstream untranslated region sequence from 28 different species, primers were designed for RT-PCR detection of tospoviruses from all recognized clades, i.e. the American, Asian and Eurasian clades, and from the small group of distinct and floating species. Pilot experiments on isolates from twenty different species showed that the designed primer sets successfully detected all species by RT-PCR, as confirmed by nucleotide sequence analysis of the amplicons. In a final optimized design, the primers were applied in a setting of five RT-PCR tests. Seven different tospoviruses were successfully identified from diagnostic samples and in addition a non-described tospovirus species from alstroemeria plants. The results demonstrate that the newly developed generic RT-PCR tests provide a relevant tool for broad detection and identification of tospoviruses in plant quarantine and diagnostic laboratories.

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