GrainGenes Reference Report: JVM-122-95
Reference
JVM-122-95
Title
Detection and relative quantitation of Soil-borne cereal mosaic virus (SBCMV) and Polymyxa graminis in winter wheat using real-time PCR (TaqMan(R))
Journal
Journal of Virological Methods
Year
2004
Volume
122
Pages
95-103
Author
[ Hide all but 1 of 6 ] Ratti C Budge G Ward L Clover G Rubies-Autonell C Henry C
Abstract
Soil-borne cereal mosaic virus (SBCMV) was first reported affecting wheat crops in Italy in 1960 and has spread subsequently to many other European countries, including the UK. SBCMV causes a serious disease of wheat, reducing yield by up to 70%; growing resistant varieties represents the only economical means of control. Real-time RT-PCR and PCR assays based on TaqMan(R) chemistry were developed for the detection and quantitation of SBCMV and its vector, Polymyxa graminis . Each assay incorporated an RNA or DNA specific internal control to facilitate quantitation. Nucleic acid extracts from SBCMV-infected plants were diluted in a nucleic acid extract from a healthy plant and amplified by real-time PCR to produce a standard curve. The standard curve was used to quantify the amount of SBCMV and P. graminis in plant samples. The sensitivity of the real-time assays were compared to established serological quantitation and conventional PCR methods by testing a range of SBCMV-infected wheat varieties. The results indicate that real-time assays were a 1000 times more sensitive than ELISA for the quantitation of SBCMV, and a 100 times more sensitive than conventional PCR for the quantitation of P. graminis . Real-time assays enabled sensitive, reproducible and specific detection of both virus and vector in wheat tissues. The real-time assays are potentially useful tools for determining variations in virus and vector concentrations in plant tissue from wheat varieties differing in resistance to SBCMV
External Databases
Pubmed: 15488626
Keyword
assays
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GrainGenes Reference Report: JVM-122-95
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GrainGenes is a product of the Agricultural Research Service of the US Department of Agriculture. | |||