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Detection of viral infections by an oligonucleotide microarray
M. Sip, D. Bystricka, S. Kmoch, L. Noskova, H. Hartmannova, P. Dedic
Jazyk angličtina Země Nizozemsko
Typ dokumentu srovnávací studie, časopisecké články, práce podpořená grantem, validační studie
- MeSH
- DNA primery MeSH
- ELISA metody MeSH
- hybridizace nukleových kyselin MeSH
- komplementární DNA genetika izolace a purifikace MeSH
- nemoci rostlin virologie MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- RNA virová genetika izolace a purifikace MeSH
- sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů metody MeSH
- Solanum tuberosum virologie MeSH
- virologie metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- validační studie MeSH
The success of DNA expression microarrays has been followed by applications of this technology to molecular diagnosis, mainly in the fields of biology and medicine. The experiments described below apply microarray diagnosis to agriculture. This report presents results of field tests for a DNA microarray designed to diagnose major viral potato pathogens. The assays were performed on samples that had been tested previously for the presence of viral infection by ELISA. RNA isolation methods were optimised for high sensitivity, using only 3 microg of total RNA that were reverse transcribed using random hexamers, with the resulting cDNA hybridised after labelling to an oligonucleotide array. The results obtained confirm the presence of pathogens indicated by ELISA and simultaneously reveal other viruses in the same reaction, showing that this method is appropriate for rapid detection of mixed viral infections. This observation was verified by subsequent RT-PCR and sequencing.
Citace poskytuje Crossref.org
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- $a Šíp, Miroslav, $d 1954- $7 xx0030656 $u Department of Laboratory Methods and Medical Technology, Faculty of Health and Social Studies, University of South Bohemia, J Boreckeho 27, 370 11 Ceske Budejovice, Czech Republic. sip@zsf.jcu.cz
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- $a The success of DNA expression microarrays has been followed by applications of this technology to molecular diagnosis, mainly in the fields of biology and medicine. The experiments described below apply microarray diagnosis to agriculture. This report presents results of field tests for a DNA microarray designed to diagnose major viral potato pathogens. The assays were performed on samples that had been tested previously for the presence of viral infection by ELISA. RNA isolation methods were optimised for high sensitivity, using only 3 microg of total RNA that were reverse transcribed using random hexamers, with the resulting cDNA hybridised after labelling to an oligonucleotide array. The results obtained confirm the presence of pathogens indicated by ELISA and simultaneously reveal other viruses in the same reaction, showing that this method is appropriate for rapid detection of mixed viral infections. This observation was verified by subsequent RT-PCR and sequencing.
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