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Development of a Reverse Transcription-Recombinase Polymerase Amplification Assay for Detection of Sugarcane Yellow Leaf Virus

文献类型： 外文期刊

作者： Feng, Xiao-Yan 1 ; Shen, Lin-Bo 1 ; Wang, Wen-Zhi 1 ; Wang, Jun-Gang 1 ; Cao, Zheng-Ying 1 ; Feng, Cui-Lian 1 ; Zhao, 1 ;

作者机构： 1.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, 4 Coll Rd, Haikou 571101, Hainan, Peoples R China

关键词： Recombinase polymerase amplification; Sugarcane; Sugarcane yellow leaf virus; Yellow leaf disease

期刊名称：SUGAR TECH （ 影响因子：1.591； 五年影响因子：1.688 ）

ISSN： 0972-1525

年卷期： 2018 年 20 卷 6 期

页码：

收录情况： SCI

摘要： Sugarcane yellow leaf virus (SCYLV, genus Polerovirus) is a harmful agent that causes sugarcane yellow leaf disease. Current methods for SCYLV detection present some limitations. In this study, a novel isothermal amplification assay, namely reverse transcription-recombinase polymerase amplification (RT-RPA), was developed to promote SCYLV detection for disease management. The assay was evaluated in terms of specificity, sensitivity, reliability, temperature limit, and time limit. The developed RT-RPA assay was highly specific and non-cross-reactive with Potato leafroll virus, which is a type of species in Polerovirus. This assay can also detect at least 10(3)-fold diluted cDNA from a SCYLV-infected sugarcane leaf, and its sensitivity is tenfold lower than that of RT-polymerase chain reaction (PCR). The reliability of the proposed assay was examined by detecting field sugarcane samples via RT-RPA and RT-PCR assays. The RT-RPA assay showed the same results as those of RT-PCR assay, indicating that the former was highly reliable for SCYLV detection. Analysis of the temperature and time limits revealed a wide operating temperature range from 27 to 45 degrees C, which was easily reached, and a rapid assay duration of 20min. In summary, the developed RT-RPA assay was rapid, specific, and reliable with acceptable sensitivity and easily reachable operating temperature and thus could be a promising tool for SCYLV diagnosis in sugarcane.

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