中文摘要：水稻齿叶矮缩病毒（RRSV）是一种非常重要的病毒，可以感染水稻，并在亚洲国家和其他主要水稻种植区造成严重的产量损失。因此，迫切需要建立一种有效、可行的现场识别和诊断方法。本研究结果表明，逆转录环介导等温扩增（RT-LAMP）反应比RT-PCR更有效和灵敏，可用于RRSV检测。最佳LAMP条件如下：0.4-1.2 muM内部引物，0.2-0.25muM外部引物，0.8μM环引物，并在62℃或63℃条件下培育30分钟。此外，RT-LAMP引物专门靶向RRSV病毒，并通过凝胶电泳和S形扩增曲线产生典型的瀑布状条带。引物不能用于扩增其他常见的植物病毒，包括番木瓜环斑病毒（PRSV）、水稻黄矮病毒（RYSV）、高粱花叶病毒（SrMV）、仙人掌X病毒X（CVX）、甜瓜黄斑病毒（MYSV）和南方水稻黑条矮缩病毒（SRBSDV）。RRSV cDNA的十倍连续稀释表明RT-LAMP在检测病毒方面比RT-PCR快得多且灵敏度至少高十倍。可以在一小时内观察到类似瀑布的产品带。在实地研究中，约77％的样本被确定为RRSV。与RT-PCR相比，RT-LAMP具有许多优点，例如低成本，高准确度、灵敏度和特异性。该技术满足现场植物病毒病快速诊断的要求，从而最好地指导种植者的管理实践。
外文摘要：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.
外文关键词：Rice ragged stunt virus (RRSV); Reverse transcription loop-mediated isothermal amplification (RT-LAMP); RT-PCR
作者：Lai, Dingwang; Zhang, Yuliang; Huang, Qixing; 等
作者单位：中国热带农业科学院
期刊名称：SAUDI JOURNAL OF BIOLOGICAL SCIENCES
期刊影响因子：2.82
出版年份：2018
出版刊次：12
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