Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (3): 517-524.doi: 10.3864/j.issn.0578-1752.2013.03.008

• PLANT PROTECTION • Previous Articles     Next Articles

A RT-LAMP Assay for Detection of Citrus Tristeza Virus

 WANG  Yong-Jiang, ZHOU  Yan, LI  Zhong-An, SU  Hua-Nan, HUANG  Ai-Jun, TANG  Ke-Zhi, ZHOU  Chang-Yong   

  1. 1.Citrus Research Institute, Chinese Academy of Agricultural Sciences, Chongqing 400712
    2.Institute of Plant Protection, Henan Academy of Agricultural Sciences, Zhengzhou 450002
    3.Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081
  • Received:2012-07-02 Online:2013-02-01 Published:2012-10-10

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Abstract: 【Objective】 The objective of this study is to establish a quick and accurate detection technique of Citrus tristeza virus by reverse transcription loop-mediated isothermal amplification (RT-LAMP). 【Method】 According to published GenBank sequences, two pairs of primers were designed in the conserved region of CTV p25. The concentration of primers, reaction time and other conditions of RT-LAMP were optimized in order to improve specificity and sensitivity.【Result】 A rapid and specific RT-LAMP method for detection of CTV was established. Sensitivity of the RT-LAMP assay was 100-fold higher than a standard RT-PCR method. The RT-LAMP products were confirmed by digestion with Acc I restriction enzyme. Products amplified by RT-LAMP were visible to add SYBR Green I, suggesting that it can be used to detect CTV directly. 【Conclusion】The detection process can be speeded up by using RT-LAMP under routine conditions at low cost and high accuracy with simple facilities and can be used as an excellent method for CTV detection in both research institutions and rural areas.

Key words: RT-LAMP , Ctrus tristeza virus (CTV) , RT-PCR , detection

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