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Development of a multiplex AmpliDet RNA for the simultaneous detection of Potato leafroll virus and Potato virus Y in potato tubers.

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.

journal_name

J Virol Methods

journal_title

Journal of virological methods

authors

Klerks MM,Leone GO,Verbeek M,van den Heuvel JF,Schoen CD

doi

10.1016/s0166-0934(01)00258-0

subject

Has Abstract

pub_date

2001-04-01 00:00:00

pages

115-25

issue

1-2

eissn

0166-0934

issn

1879-0984

pii

S0166093401002580

journal_volume

93

pub_type

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