Simplified sample preparation for detection of wheat streak mosaic virus and barley yellow dwarf virus by PCR.
A PCR diagnostic procedure for wheat streak mosaic virus (WSMV) was developed using a primer derived from 3'-terminal sequences of five WSMV isolates and an oligo d(T)-based primer. Cereal extracts prepared by digestion with proteinase K and boiling permitted PCR-based detection of both WSMV and BYDV in field samples. This procedure saves time, eliminates multiple liquid transfer steps, and reduces the chances of cross contamination. Sensitivity of such assays is still very good; BYDV could be readily detected in plant sap diluted over 1000-fold. Further, parallel detection of WSMV and barley yellow dwarf virus (BYDV) in the same samples is possible with this method.
French
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United States Department of Agriculture-Agricultural Research Service, University of Nebraska, Lincoln 68583.
Robertson
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NL
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D005069:Evaluation Studies as Topic; D017793:Luteovirus / Q000235:genetics* / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D014773:Virology / Q000379:methods
D017931:DNA Primers; D004279:DNA, Viral