| 李属坏死环斑病毒（PNRSV）RT-LAMP检测方法的建立 |
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| 引用本文： | 韩剑,;罗明,;殷智婷,;周国辉,;张祥林. 李属坏死环斑病毒（PNRSV）RT-LAMP检测方法的建立[J]. 新疆农业大学学报, 2014, 0(4): 327-332 |
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| 作者姓名： | 韩剑, 罗明, 殷智婷, 周国辉, 张祥林 |
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| 作者单位： | [1]新疆农业大学农学院,乌鲁木齐830052; [2]华南农业大学资源与环境学院,广州510642; [3]新疆出入境检疫检验局,乌鲁木齐830052 |
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| 基金项目： | 新疆维吾尔自治区自然科学基金项目（2012211B25） |
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| 摘 要： | 针对李属坏死环斑病毒（PNRSV）外壳蛋白（CP）基因保守序列,设计了1套特异性识别CP基因中6个不同区段的环介导等温扩增（LAMP）引物。通过对反应条件的优化,建立了适用于PNRSV的RT-LAMP检测技术,对优化后的方法进行特异性、灵敏度评价。结果显示,此方法能够特异性的检测出PNRSV,对马铃薯X病毒（PVX）、马铃薯Y病毒（PVY）、黄瓜花叶病毒（CMV）、蚕豆萎蔫病毒（BBWV）的检测均为阴性,灵敏度高于RT-PCR 10倍。
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| 关 键 词： | 李属坏死环斑病毒(PNRSV) 反转录环介导等温扩增(RT-LAMP) CP基因 检测 |
Development of Reverse Transcription Loop-mediated Isothermal Amplification Assay for Detection of Prunus Necrotic Ringspot Virus |
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| Affiliation: | HAN Jian, LUO Ming , YIN Zhi-ting , ZHOU Guo-hui, ZHANG Xiang-lin (1.College of Agronomy, Xinjiang Agricultural University, Urumqi 830052, China; 2. College of Natural Resources and Environment, South China Agricultural University, Guangzhou 510642, China; 3.Xinjiang Entry and Exit Inspection and Quarantine Bureau,Urumqi 830052,China) |
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| Abstract: | A set of four specific LAMP primers were designed to recognize six distinct regions,according to the gene sequence of coat protein in prunus necrotic ringspot virus（ PNRSV）. Based on optimization of reaction conditions,RT-LAMP detection method was developed which was treated with specific techniques and was evaluated. It suggested that PNRSV can be found by the method. The amplified fragments were not found in the samples of potato virus X（ PVX）,potato virus Y（ PVY）,cucumber mosaic virus（ CMV）,broad bean wilt virus（ BBWV） and the sensitivity for the detection of PNRSV was 10 times higher than the traditional RT-PCR method. |
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