文章摘要
李战彪,谢慧婷,崔丽贤,蔡健和,秦碧霞.甘蔗线条花叶病毒 RT-LAMP 检测方法的建立及应用(英文)[J].广东农业科学,2020,47(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)
Establishment and Application of RT-LAMP Detection Method for Sugarcane Streak Mosaic Virus
  
DOI:10.16768/j.issn.1004-874X.2020.04.013
中文关键词: :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
英文关键词: sugarcane  sugarcane streak mosaic virus(SCSMV)  reverse transcription loop-mediated isothermal amplification(RT-LAMP)  coat protein gene  field detection
基金项目:Major Science and Technology Project of Guangxi Province(Guike AA17202042-12);Basic Research Project of Guangxi Academy of Agricultural Sciences(2015YT42)
作者单位
李战彪,谢慧婷,崔丽贤,蔡健和,秦碧霞 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广西 南宁 530007 
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中文摘要:
      【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。
英文摘要:
      【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 RTLAMP 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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