Development of a Reverse Transcription-Recombinase Polymerase Amplification Assay for Detection of Sugarcane Yellow Leaf Virus
Development of a Reverse Transcription-Recombinase Polymerase Amplification Assay for Detection of Sugarcane Yellow Leaf Virus
复制标题
开发用于检测甘蔗黄叶病毒的逆转录重组酶聚合酶扩增测定法
DOI：
10.1007/s12355-018-0602-6
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发表时间：
2018
期刊：
影响因子：
1.9
通讯作者：
Zhang Shu-Zhen
中科院分区：
文献类型：
--
作者：
Feng Xiao-Yan;Shen Lin-Bo;Wang Wen-Zhi;Wang Jun-Gang;Cao Zheng-Ying;Feng Cui-Lian;Zhao Ting-Ting;Zhang Shu-Zhen
Sugarcane yellow leaf virus(SCYLV, genusPolerovirus) 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 withPotato leafroll virus, which is a type of species inPolerovirus. This assay can also detect at least 103-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 °C, which was easily reached, and a rapid assay duration of 20 min. 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.
