Abstract：
:A real-time TaqMan RT-PCR assay was developed for the rapid and sensitive detection of Tomato ringspot virus (ToRSV), an important plant virus which infects a wide range of fruit and ornamental crops. Primers and a probe were designed based on the highly conserved 3'-untranslated region (UTR) sequences of ToRSV, to amplify a 182bp fragment of this region of RNA-1 and RNA-2. The assay was demonstrated to reliably amplify all ToRSV isolates tested. The detection limit was estimated to be about 12 copies of the ToRSV target region. No amplification was observed from the RNA of other nepoviruses or healthy host species. A comparison with a published conventional RT-PCR and a SYBR-based qRT-PCR indicated that both of the published assays lacked reliability and sensitivity, as neither were able to amplify all ToRSV isolates tested, and both were approximately 1000 times less sensitive than the novel TaqMan real-time assay. This TaqMan real-time assay was tested using four different reagent kits and was shown to be robust and stable, with no significant differences in sensitivity between kits. It is expected that the implementation of this TaqMan real-time RT-PCR assay will facilitate efficient phytosanitary certification of nursery stock requiring testing for ToRSV by regulatory agencies, and will also have wider uses for the general detection of ToRSV in a range of hosts.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Tang J,Khan S,Delmiglio C,Ward LIdoi
10.1016/j.jviromet.2014.02.011subject
Has Abstractpub_date
2014-06-01 00:00:00pages
38-43eissn
0166-0934issn
1879-0984pii
S0166-0934(14)00054-8journal_volume
201pub_type
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journal_title：Journal of virological methods
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pub_type： 临床试验,杂志文章
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(98)00054-8
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journal_title：Journal of virological methods
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journal_title：Journal of virological methods
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journal_title：Journal of virological methods
pub_type： 杂志文章
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journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2004.08.012
更新日期：2004-12-01 00:00:00
abstract：:The performance of the rapid confirmatory HIV 1/2 assay Geenius was compared with the conventional HIV 1/2 line immunoblot (INNO-LIA HIV I/II Score). One hundred HIV 1/2 confirmed positive samples from donors and patients and 136 negative screening samples from blood donors were evaluated with both assays. A 20 member...
journal_title：Journal of virological methods
pub_type： 杂志文章
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更新日期：2014-09-01 00:00:00
