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当前位置: 首页 > 文章 > 甘蔗线条花叶病毒RT-LAMP检测方法的建立及应用（英文） 广东农业科学 2020 (4) 92-98
Position: Home > Articles > Establishment and Application of RT-LAMP Detection Method for Sugarcane Streak Mosaic Virus Guangdong Agricultural Sciences 2020 (4) 92-98

甘蔗线条花叶病毒RT-LAMP检测方法的建立及应用（英文）

作  者：
李战彪;谢慧婷;崔丽贤;蔡健和;秦碧霞
单  位：
广西农业科学院植物保护研究所/广西作物病虫害生物学重点实验室
关键词：
甘蔗;甘蔗线条花叶病毒(SCSMV);环介导等温扩增(RT-LAMP);外壳蛋白基因;田间检测
摘  要：
【目的】建立甘蔗线条花叶病毒(Sugarcane streak mosaic virus,SCSMV)的RT-LAMP检测方法,为SCSMV的检测提供技术支持。【方法】根据甘蔗线条花叶病毒外壳蛋白基因的保守序列设计特异引物,优化RT-LAMP的反应条件,对比分析RT-LAMP的检测灵敏度和特异性;并在RT-LAMP反应产物中加入SYBR GREEN I进行可视化检测,方便快速判定结果。【结果】成功建立了一种用于检测SCSMV的RT-LAMP方法,可在60 ～ 63 ℃下60 min完成对SCSMV的检测,且具有较高的灵敏度,比传统RT-PCR高100 倍,最低检测RNA浓度为167.9×10~(-5) ng。此外,该方法具有较高的特异性,可特异检测SCSMV。【结论】建立了一种灵敏、简便、快速的SCSMV RT-LAMP检测方法并用于田间甘蔗样品的检测。
译  名：
Establishment and Application of RT-LAMP Detection Method for Sugarcane Streak Mosaic Virus
作  者：
LI Zhanbiao;XIE Huiting;CUI Lixian;CAI Jianhe;QIN Bixia;Institute of Plant Protection, Guangxi Academy of Agricultural Sciences, Guangxi Key Laboratory of Biology for Crop Diseases and Insect Pests;
关键词：
sugarcane;;sugarcane streak mosaic virus(SCSMV);;reverse transcription loop-mediated isothermal amplification(RT-LAMP);;coat protein gene;;field detection
摘  要：
【Objective】In order to provide technical support for the detection of sugarcane streak mosaic virus(SCSMV),a reverse transcription loop-mediated isothermal amplification(RT-LAMP)assay was established.【Method】A set of specific primers were designed from the conserved regions of coat protein gene sequences of SCSMV.The reaction conditions for RT-LAMP were optimized and the detection sensibility and specificity of RT-LAMP were analyzed and compared.For fluorescence detection,the RT-LAMP reaction products were also visually detected by adding SYBR Green I dye to identify the results in a convenient and rapid way.【Result】A RT-LAMP assay was developed for the rapid detection of SCSMV,and the whole detection process could be completed in 60 minutes at 60-63℃.The RT-LAMP established in this study had high sensitivity,which was 100 times higher than that of the traditional RT-PCR,and the lowest detection concentration was 167.9×10~(-5) ng.In addition,the method had high specificity and could be used for the specific detection of SCSMV.【Conclusion】A sensitive,convenient and rapid RT-LAMP assay was established and used for field detection of SCSMV samples.

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