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当前位置: 首页 > 文章 > 马铃薯S病毒PVS~O株系RT-LAMP检测方法的建立及应用 园艺学报 2018 (8) 1613-1620
Position: Home > Articles > Establishment of Loop-mediated Isothermal Amplification Assay for Detection of Potato virus S Ordinary Strain Acta Horticulturae Sinica 2018 (8) 1613-1620

马铃薯S病毒PVS~O株系RT-LAMP检测方法的建立及应用

作  者：
李华伟;许泳清;罗文彬;纪荣昌;刘中华;许国春;张鸿;李国良;林赵淼;邱永祥;邱思鑫;汤浩
关键词：
马铃薯;马铃薯S病毒;环介导等温扩增;钙黄绿素
摘  要：
为建立一种马铃薯S病毒普通株系PVS~O快速、灵敏的RT-LAMP可视化检测方法。根据马铃薯S病毒普通株系的CP核苷酸序列设计4条引物,以感染PVS的马铃薯叶片总RNA为模板,采用一步法RT-LAMP,在62℃下反应60 min,扩增产物利用琼脂糖电泳分析和钙黄绿素染料显色判断结果,同时对其特异性和灵敏性进行测定。结果表明,建立的RT-LAMP方法可特异地对PVS~O进行检测,检测灵敏度为RT-PCR的100倍。对33份马铃薯样品检测表明,RT-LAMP与RT-PCR方法检测结果基本一致。因此建立的PVS~O的RT-LAMP可视化检测方法简便,特异性强,灵敏度高,适合PVS~O的快速检测。
译  名：
Establishment of Loop-mediated Isothermal Amplification Assay for Detection of Potato virus S Ordinary Strain
作  者：
LI Huawei;XU Yongqing;LUO Wenbin;JI Rongchang;LIU Zhonghua;XU Guochun;ZHANG Hong;LI Guoliang;LIN Zhaomiao;QIU Yongxiang;QIU Sixin;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;
单  位：
LI Huawei%XU Yongqing%LUO Wenbin%JI Rongchang%LIU Zhonghua%XU Guochun%ZHANG Hong%LI Guoliang%LIN Zhaomiao%QIU Yongxiang%QIU Sixin%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
关键词：
potato;;Potato virus S;;LAMP;;calcein
摘  要：
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(PVS~O). Four RT-LAMP primers were designed on the basis of the coat protein(CP)gene sequences of PVS~O,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 PVS~O 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 PVS~O.

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