Articles
THE IMPROVED DETECTION OF STRAWBERRY CRINKLE VIRUS USING REAL-TIME RT-PCR (TAQMAN®)
Article number
656_11
Pages
81 – 86
Language
English
Abstract
Strawberry crinkle virus (SCV; Genus Cytorhabdovirus) is a widespread and economically important quarantine pathogen of Fragaria spp.
The development of accurate and reliable diagnostic methods for the detection of SCV has been hampered severely, as the virus is hard to purify and as strawberry is a difficult host for RNA extraction.
As a result of these factors, an alternative approach using real-time RT-PCR (TaqMan) for the routine detection of SCV was investigated.
A SCV-specific TaqMan assay was developed and tested against a range of UK isolates, including one shown to have 10% sequence divergence from the consensus sequence found in other isolates.
The assay has also detected SCV from a number of field-grown strawberry samples.
The overall efficiency and reliability of the assay was improved through the use of an internal control assay and the development of methods that allow reliable RNA extraction from strawberry.
The performance of the new test was compared to conventional and nested RT-PCR.
The development of accurate and reliable diagnostic methods for the detection of SCV has been hampered severely, as the virus is hard to purify and as strawberry is a difficult host for RNA extraction.
As a result of these factors, an alternative approach using real-time RT-PCR (TaqMan) for the routine detection of SCV was investigated.
A SCV-specific TaqMan assay was developed and tested against a range of UK isolates, including one shown to have 10% sequence divergence from the consensus sequence found in other isolates.
The assay has also detected SCV from a number of field-grown strawberry samples.
The overall efficiency and reliability of the assay was improved through the use of an internal control assay and the development of methods that allow reliable RNA extraction from strawberry.
The performance of the new test was compared to conventional and nested RT-PCR.
Authors
R.A. Mumford, A.L. Skelton, N. Boonham, K.I. Posthuma, M.J. Kirby, A.N. Adams
Keywords
Diagnosis, Fragaria, cytorhabdovirus, Rhabdoviridae
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