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Loop-mediated isothermal amplification for rapid and precise detection of the burrowing nematode, Radopholus similis , directly from diseased plant tissues

Loop-mediated isothermal amplification for rapid and precise detection of the burrowing nematode,... A novel, simple, rapid and highly sensitive assay and diagnostic tool for the burrowing nematode, Radopholus similis , was developed using a loop-mediated isothermal amplification (LAMP). The LAMP assay was targeted on the specific fragments of rRNA gene D2-D3 regions of R. similis . The detection limitation of the LAMP assay was as low as ten copies of plasmid DNA containing the target DNA, 10 fg of genomic DNA and 5 × 10 −5 nematodes. The detection sensitivity of the LAMP method for R. similis DNA was 10-100 times higher than normal PCR-based detection methods. The LAMP amplifications could be observed directly by eye by adding SYBR Green I and the lateral flow dipstick (LFD). LAMP assay for R. similis is a practical and useful diagnostic tool for early diagnosis of plant tissues infested by R. similis . The LAMP assay developed in this study is highly effective, easy to perform and readily adaptable for diagnostic and monitoring of the R. similis -diseased seedling in the field. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Nematology Brill

Loop-mediated isothermal amplification for rapid and precise detection of the burrowing nematode, Radopholus similis , directly from diseased plant tissues

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Publisher
Brill
Copyright
© Koninklijke Brill NV, Leiden, The Netherlands
Subject
Articles
ISSN
1388-5545
eISSN
1568-5411
DOI
10.1163/156854112X638415
Publisher site
See Article on Publisher Site

Abstract

A novel, simple, rapid and highly sensitive assay and diagnostic tool for the burrowing nematode, Radopholus similis , was developed using a loop-mediated isothermal amplification (LAMP). The LAMP assay was targeted on the specific fragments of rRNA gene D2-D3 regions of R. similis . The detection limitation of the LAMP assay was as low as ten copies of plasmid DNA containing the target DNA, 10 fg of genomic DNA and 5 × 10 −5 nematodes. The detection sensitivity of the LAMP method for R. similis DNA was 10-100 times higher than normal PCR-based detection methods. The LAMP amplifications could be observed directly by eye by adding SYBR Green I and the lateral flow dipstick (LFD). LAMP assay for R. similis is a practical and useful diagnostic tool for early diagnosis of plant tissues infested by R. similis . The LAMP assay developed in this study is highly effective, easy to perform and readily adaptable for diagnostic and monitoring of the R. similis -diseased seedling in the field.

Journal

NematologyBrill

Published: Jan 1, 2012

Keywords: D2-D3; diagnostic test; molecular assay; nematode detection

References