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dc.contributor.authorKorimbocus, J.*
dc.contributor.authorCoates, David*
dc.contributor.authorBarker, I.*
dc.contributor.authorBoonham, N.*
dc.date.accessioned2009-12-07T15:55:50Z
dc.date.available2009-12-07T15:55:50Z
dc.date.issued2002
dc.identifier.citationKorimbocus, J., Coates, D., Barker, I. and Boonham, N. (2002). Improved detection of Sugarcane yellow leaf virus using a real- time fluorescent (TaqMan) RT-PCR assay. Journal of Virological Methods, Vol. 103, No. 2, pp.109-120.en
dc.identifier.urihttp://hdl.handle.net/10454/4037
dc.descriptionnoen
dc.description.abstractYellow leaf syndrome (YLS) of sugarcane has been associated with Sugarcane yellow leaf virus (ScYLV) and has been reported from most sugarcane growing countries around the world. As sugarcane is vegetatively propagated, it is important to use effective and sensitive detection methods to screen new propagating material. Virus detection in symptomatic tissue is currently achieved using enzyme linked immunosorbent assay (ELISA), tissue blot immunoassay (TBIA) or a conventional RT-PCR based assay. This paper reports the development of an improved assay based on multiplex real-time fluorescent RT-PCR. The new assay is 100-fold more sensitive than conventional RT-PCR, and incorporates a novel `RNA specific¿ internal positive control (based around the intron of the caffeic acid 3-o-methyltransferase gene) to guard against false negative results. The paper also describes the comparison of eight RNA extraction methods for sugarcane tissue giving a number of alternatives for different laboratory situations. The sensitivity of this assay has allowed the detection of ScYLV in many samples that were thought to be healthy following conventional testing (RT-PCR, ELISA or TBIA). The detection of ScYLV using this TaqMan assay can be applied to the production of ScYLV-free plants and prevents its spread through the propagation material.en
dc.language.isoenen
dc.publisherElsevieren
dc.relation.isreferencedbyhttp://dx.doi.org/10.1016/S0166-0934(01)00406-2en
dc.subjectSugarcane yellow leaf virusen
dc.subjectReal-time fluorescent TaqManen
dc.subjectRT-PCRen
dc.subjectVirus detectionen
dc.subjectLuteoviridaeen
dc.titleImproved detection of Sugarcane yellow leaf virus using a real- time fluorescent (TaqMan) RT-PCR assay.en
dc.status.refereedYesen
dc.typeArticleen
dc.type.versionpublished version paperen


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