4.4 Article

Rapid detection of Prunus necrotic ringspot virus using magnetic nanoparticle-assisted reverse transcription loop-mediated isothermal amplification

Journal

JOURNAL OF VIROLOGICAL METHODS
Volume 208, Issue -, Pages 85-89

Publisher

ELSEVIER
DOI: 10.1016/j.jviromet.2014.07.033

Keywords

PNRSV; RT-LAMP; Detection

Funding

  1. Special Fund for Agro-scientific Research in the Public Interest [201203075]
  2. National Key Technology Research and Development Program of the Ministry of Science and Technology of China [2013BAD02B03-3-2]
  3. Special Fund for Innovation Teams of Fruit trees in Agricultural Technology System of Shandong Province [SDAIT-03-022-04]

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Prunus necrotic ringspot virus (PNRSV) has seriously reduced the yield of Prunus species worldwide. In this study, a highly efficient and specific two-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed to detect PNRSV. Total RNA was extracted from sweet cherry leaf samples using a commercial kit based on a magnetic nanoparticle technique. Transcripts were used as the templates for the assay. The results of this assay can be detected using agarose gel electrophoresis or by assessing in-tube fluorescence after adding SYBR Green I. The assay is highly specific for PNRSV, and it is more sensitive than reverse-transcription polymerase chain reaction (RT-PCR). Restriction enzyme digestion verified further the reliability of this RT-LAMP assay. To our knowledge, this is the first report of the application of RT-LAMP to PNRSV detection in Prunus species. (C) 2014 Elsevier B.V. All rights reserved.

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