Establishment of Loop-mediated Isothermal Amplification Assay for Detection of Potato virus S Ordinary Strain

doi:10.16420/j.issn.0513-353x.2018-0164

ACTA HORTICULTURAE SINICA ›› 2018, Vol. 45 ›› Issue (8): 1613-1620.doi: 10.16420/j.issn.0513-353x.2018-0164

• New Technologies and New Methods • Previous Articles Next Articles

Establishment of Loop-mediated Isothermal Amplification Assay for Detection of Potato virus S Ordinary Strain

LI Huawei，XU Yongqing，LUO Wenbin，JI Rongchang，LIU Zhonghua，XU Guochun，ZHANG Hong，LI Guoliang，LIN Zhaomiao，QIU Yongxiang，QIU Sixin，and TANG Hao*

Crop Research Institute，Fujian Academy of Agricultural Sciences，Scientific Observing and Experimental Station of Tuber and Root Crops in South China，Ministry of Agricultrue，P. R. China，Fuzhou 350013，China

Online:2018-08-25 Published:2018-08-25

Abstract

Abstract:

The object of this study is to develop a One-step reverse transcription loop-mediated isothermal amplification（RT-LAMP）assay for rapid and sensitive detection of Potato virus S ordinary strain（PVSO）. Four RT-LAMP primers were designed on the basis of the coat protein（CP）gene sequences of PVSO，and template RNA from infected leaves were used for One-step RT-LAMP which were carried out under isothermal conditions at 62 ℃ for 60 minutes. RT-LAMP products were analyzed by electrophoresis in agarose gels followed by staining with Calcein. Then the specificity and sensitivity of RT-LAMP were testified. A rapid and specific RT-LAMP method for detection of PVSO was established. Sensitivity of the RT-LAMP assay was 100-fold higher than ordinary RT-PCR method，and the testing results of 33 potato samples were basically consistent with RT-PCR. The RT-LAMP described in this study represents a sensitive，specific and rapid assay for the detection of PVSO.

Key words: potato, Potato virus S, LAMP, calcein

CLC Number:

S 641

LI Huawei，XU Yongqing，LUO Wenbin，JI Rongchang，LIU Zhonghua，XU Guochun，ZHANG Hong，LI Guoliang，LIN Zhaomiao，QIU Yongxiang，QIU Sixin，and TANG Hao*. Establishment of Loop-mediated Isothermal Amplification Assay for Detection of Potato virus S Ordinary Strain[J]. ACTA HORTICULTURAE SINICA, 2018, 45(8): 1613-1620.

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Establishment of Loop-mediated Isothermal Amplification Assay for Detection of Potato virus S Ordinary Strain

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