Abstract
A novel isothermal multiplex AmpliDet RNA system is described for the simultaneous amplification and detection of Potato leafroll virus (PLRV) and Potato virus Y (PVY) in seed potatoes. The risk of contamination by carry-over during diagnostic screening is eliminated by performing the reaction in a single closed tube. The viruses present in a sample are identified using differently coloured molecular beacons directed to a selected virus-specific sequence within the amplicon formed during amplification. With this system, as little as 10 fg of purified PLRV or PVY can be detected. The presence of both viruses in a sample is detected by the multiplex assay within a high range of virus concentrations. The reliability of the multiplex assay was compared with the enzyme-linked immunosorbent assay for detection of PLRV- or PVY-antigens in potato tubers. The multiplex assay detected clearly the viruses present originally in the potato tubers in all samples, demonstrating its potential for routine diagnostic work and high-throughput screening.
Original language | English |
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Pages (from-to) | 115-125 |
Journal | Journal of Virological Methods |
Volume | 93 |
Issue number | 1-2 |
DOIs | |
Publication status | Published - 2001 |
Keywords
- AmpliDet RNA
- Fluorescent probes
- Homogeneous assays
- Molecular beacons
- Multiplex detection
- NASBA
- Potato viruses