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

文献类型: 外文期刊

作者: Peng, Jun 1 ; Zhang, Junfang 3 ; Xia, Zihao 1 ; Li, Yongqiang 1 ; Huang, Junsheng 4 ; Fan, Zaifeng 1 ;

作者机构: 1.China Agr Univ, State Key Lab Agrobiotechnol, Beijing 100193, Peoples R China

2.China Agr Univ, Minist Agr, Key Lab Plant Pathol, Beijing 100193, Peoples R China

3.Chinese Acad Trop Agr Sci, Environm & Plant Protect Inst, Key Lab Monitoring & Control Trop Agr & Forest In, Danzhou 571737, Hainan, Peoples R China

4.Chinese Acad Trop Agr Sci, Environm & Plant Protect Inst, Key Lab Monitoring

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

期刊名称:JOURNAL OF VIROLOGICAL METHODS ( 影响因子:2.014; 五年影响因子:2.001 )

ISSN: 0166-0934

年卷期: 2012 年 185 卷 2 期

页码:

收录情况: SCI

摘要: 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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