DNA microarray: parallel detection of potato viruses.
DNA microarray assay has become a useful tool for gene expression studies. Less frequent is its application to detection of viruses or diagnostics of virus diseases. Here we show design of a microscope slide-based microarray assay for simultaneous identification of several potato viruses. Different primer pairs were designed or adopted to obtain specific amplicons from six potato viruses: Potato virus A (PVA), Potato virus S (PVS), Potato virus X (PVX), Potato virus Y (PVY), Potato mop-top virus (PMTV) and Potato leaf-roll virus (PLRV). Purified viral DNA probes were spotted on a microscope slide coated with poly-L-lysine. The same primers were used for preparation of fluorochrome-labeled targets. The latter were denatured and hybridized on the microarray slide (chip). An example of simultaneous assay of two pathogens is given and possibilities of practical application of this type of assay are discussed.
Bystrická
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Department of Plant Virology, Institute of Plant Molecular Biology, Academy of Sciences of the Czech Republic, Branisovská 31, 370 05 Ceské Budĕjovice, Czech Republic.
Lenz
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Mráz
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Dĕdic
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Síp
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D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods* / Q000706:statistics & numerical data; D010935:Plant Diseases / Q000821:virology*; D010942:Plant Viruses / Q000235:genetics / Q000302:isolation & purification*; D017863:Potexvirus / Q000235:genetics / Q000302:isolation & purification; D017800:Potyvirus / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D011198:Solanum tuberosum / Q000821:virology*
D017931:DNA Primers; D004279:DNA, Viral