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

被引：36

作者：

Klerks, MM ^{[1
]}

Leone, GOM ^{[1
]}

Verbeek, M ^{[1
]}

van den Heuvel, JFJM ^{[1
]}

Schoen, CD ^{[1
]}

机构：

[1] Plant Res Int BV, NL-6700 AA Wageningen, Netherlands

来源：

JOURNAL OF VIROLOGICAL METHODS | 2001年 / 93卷 / 1-2期

关键词：

AmpliDet RNA; fluorescent probes; homogeneous assays; molecular beacons; multiplex detection; NASBA; potato viruses;

D O I：

10.1016/S0166-0934(01)00258-0

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. (C) 2001 Elsevier Science B.V. All rights reserved.

引用

页码：115 / 125

页数：11

共 19 条

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