TY - JOUR
T1 - Generic RT-PCR tests for detection and identification of tospoviruses
AU - Hassani-Mehraban, A.
AU - Westenberg, M.
AU - Verhoeven, J.T.J.
AU - van de Vossenberg, B.T.L.H.
AU - Kormelink, R.
AU - Roenhorst, J.W.
PY - 2016
Y1 - 2016
N2 - 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.
AB - 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.
KW - Bunyaviridae
KW - Clade-specific primers
KW - Diagnostics
KW - Nucleocapsid gene
KW - Plant virus
U2 - 10.1016/j.jviromet.2016.03.015
DO - 10.1016/j.jviromet.2016.03.015
M3 - Article
AN - SCOPUS:84962339701
SN - 0166-0934
VL - 233
SP - 89
EP - 96
JO - Journal of Virological Methods
JF - Journal of Virological Methods
ER -