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| 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 |
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| Abstract: |
| 【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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