4.4 Article

Rapid and sensitive detection of Banana bunchy top virus by loop-mediated isothermal amplification

Journal

JOURNAL OF VIROLOGICAL METHODS
Volume 185, Issue 2, Pages 254-258

Publisher

ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2012.06.026

Keywords

Banana bunchy top virus; Virus detection; Loop-mediated isothermal amplification (LAMP); Turbidimetry

Funding

  1. Special Fund for Agro-Scientific Research in the Public Interest [200903049]
  2. Ministry of Education [IRT1042]
  3. Research Fund for the Doctoral Program of Higher Education of China [20110008110020]
  4. Chinese National Non-profit Institute Research Grant of CATAS-EPI [2009hzs1J07]

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A sensitive loop-mediated isothermal amplification (LAMP) assay was developed for rapid detection of Banana bunchy top virus (BBTV) infection. The reaction was performed in a single tube at 63 degrees C for 90 mm, with an improved closed-tube detection system by adding the SYBR Green I dye to the inside of the tube lid prior to amplification. The detection limit of the LAMP assay was approximately 1 pg/mu l plasmid DNA when mixed with extracted DNA from healthy banana plant, and no cross-reaction with other banana-infected pathogens was observed. Real-time turbidimetry was used to monitor the amplification result in the tubes, and it was shown that this LAMP assay was about 100-fold more sensitive than PCR. The results demonstrated that this LAMP method should be useful for both banana disease monitoring and mass propagation of virus-free banana plantlets. (C) 2012 Elsevier B.V. All rights reserved.

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