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Improved detection of Sugarcane yellow leaf virus using a real- time fluorescent (TaqMan) RT-PCR assay.

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dc.creator Korimbocus, J.
dc.creator Coates, David
dc.creator Barker, I.
dc.creator Boonham, N.
dc.date 2009-12-07T15:55:50Z
dc.date 2009-12-07T15:55:50Z
dc.date 2002
dc.identifier Korimbocus, 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.
dc.identifier http://hdl.handle.net/10454/4037
dc.description no
dc.description Yellow 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.
dc.language en
dc.publisher Elsevier
dc.relation http://dx.doi.org/10.1016/S0166-0934(01)00406-2
dc.subject Sugarcane yellow leaf virus
dc.subject Real-time fluorescent TaqMan
dc.subject RT-PCR
dc.subject Virus detection
dc.subject Luteoviridae
dc.title Improved detection of Sugarcane yellow leaf virus using a real- time fluorescent (TaqMan) RT-PCR assay.
dc.type Article
dc.type published version paper


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