Reverse transcription loop-mediated isothermal amplification to rapidly detect Rice ragged stunt virus

文献类型： 外文期刊

第一作者： Lai, Dingwang

作者： Lai, Dingwang;Zhang, Yuliang;Huang, Qixing;Yin, Guohua;Liu, Zhixin;Guo, Anping;Lai, Dingwang;Yin, Guohua;Pennerman, Kayla K.

作者机构：

关键词： Rice ragged stunt virus (RRSV); Reverse transcription loop-mediated isothermal amplification (RT-LAMP); RT-PCR

期刊名称：SAUDI JOURNAL OF BIOLOGICAL SCIENCES （ 影响因子：4.219； 五年影响因子：4.432 ）

ISSN： 1319-562X

年卷期： 2018 年 25 卷 8 期

页码：

收录情况： SCI

摘要： Rice ragged stunt virus (RRSV) is a very important virus that infects rice and causes serious yield losses in Asian countries and other major rice planting areas. Thus, it is urgent to establish an efficient and practical approach for identification and diagnosis in the field. Our results indicated that reverse transcription loop-mediated isothermal amplification (RT-LAMP) reactions are more efficient and sensitive than RT-PCR for RRSV detection. The optimal LAMP conditions were as follows: 0.4-1.2 mu M internal primers, 0.2-0.25 mu M external primers, 0.8 mu M loop primers, and incubation at 62 degrees C or 63 degrees C for 30 min. Furthermore, the RT-LAMP primers specifically targeted RRSV virus and resulted in typical waterfall-like bands by gel electrophoresis and sigmoidal amplification curves. The primers could not be used to amplify other common plant viruses including Papaya ringspot virus (PRSV), Rice yellow stunt virus (RYSV), Sorghum mosaic virus (SrMV), Cactus virus X (CVX), Melon yellow spot virus (MYSV) and Southern rice black-streaked dwarf virus (SRBSDV). Ten-fold serial dilutions of RRSV cDNA indicated that RT-LAMP is much faster and at least ten times more sensitive than RT-PCR in detecting the virus. The waterfall-like product bands could be observed within one hour. In the field study, about 77% samples were identified as RRSV. RT-LAMP has many benefits over RT-PCR such as low cost and high accuracy, sensitivity, and specificity. This technology meets the requirements for rapid diagnosis of plant virus diseases in the field to best guide management practices for growers. (C) 2016 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University.

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