| 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 |
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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 |
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| 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） |
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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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