Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (18): 3939-3945.doi: 10.3864/j.issn.0578-1752.2013.18.025

• RESEARCH NOTES • Previous Articles     Next Articles

Detection of Sweet potato chlorotic stunt virus Strain WA by Reverse Transcription Loop-Mediated Isothermal Ampli?cation Reaction

 QIAO  Qi, ZHANG  Zhen-Chen, QIN  Yan-Hong, ZHANG  De-Sheng, TIAN  Yu-Ting, WANG  Shuang, WANG  Yong-Jiang   

  1. Institute of Plant Protection, Henan Academy of Agricultural Sciences/Henan Key Laboratory of Crop Pest Control/IPM Key Laboratory in Southern Part of North China for Ministry of Agriculture, Zhengzhou 450002
  • Received:2013-03-26 Online:2013-09-15 Published:2013-06-17

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Abstract: 【Objective】The objective of this study is to develop a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid and sensitive detection of Sweet potato chlorotic stunt virus strain WA (SPCSV-WA). 【Method】Four RT-LAMP primers, designed on the basis of the coat protein (CP) gene sequences of SPCSV-WA, and template RNA from infected leaves were used for one-step RT-LAMP which were carried out under isothermal conditions at 65℃ for 1 h. RT-LAMP products were analyzed by electrophoresis in agarose gels followed by staining with SYBR green I. A fragment from the RT-LAMP product was then cloned and sequenced to con?rm the spect?city of the assay. The RT-LAMP, and meanwhile compared with the conventional RT-PCR assay, was performed to detect SPCSV-WA from 14 sweet potato samples. 【Result】The RT-LAMP showed a high specificity and the detection limit was 101 copies/µL RNA template. A fragment cloned from the RT-LAMP product indicated that the nucleotide sequence cloned was SPCSV-WA CP. Fourteen sweet potato samples testing results of RT-LAMP were consistent with that of RT-PCR. 【Conclusion】 The RT-LAMP described in this study represents a very sensitive, speci?c and rapid assay for the detection of SPCSV-WA.

Key words: Ipomoea batatas , Sweet potato chlorotic stunt virus strain WA , RT-LAMP , detection

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