| 实时荧光RT-PCR一步法检测番茄环斑病毒 |
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| 引用本文： | 朱建裕,朱水芳,廖晓兰,高必达.实时荧光RT-PCR一步法检测番茄环斑病毒[J].植物病理学报,2003,33(4):338-341. |
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| 作者姓名： | 朱建裕 朱水芳 廖晓兰 高必达 |
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| 作者单位： | 1 湖南农业大学植保系, 长沙 410128;2国家出入境检验检疫局植检所, 北京 100029 |
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| 摘 要： | 根据番茄环斑病毒(ToRSV)各株系RNA1聚合酶基因的保守序列,设计并合成1对引物和1条TaqMan荧光探针,建立了对ToRSV的实时荧光RT-PCR检测方法。该方法利用TaqMan荧光探针实时监测目的基因的扩增,实现RT-PCR扩增与探针检测同时进行,不需PCR后处理,消除了PCR产物的污染,不需电泳和EB染色,减少了检测步骤,节省了时间。这个方法具有"灵敏、准确、简便、快速"的特点适合于种传病害的快速诊断和口岸检验检疫运用。
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| 关 键 词： | 番茄环斑病毒(ToRSV) 种传病害 实时荧光RT-PCR 检测 |
| 文章编号： | 0412-0914(2003)04-0338-04 |
| 修稿时间： | 2002年11月2日 |
Detection of Tomato ring spot virus by real-time fluorescent RT-PCR one step assay |
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| ZHU Jian yu ,ZHU Shui fang ,LIAO Xiao lan ,GAO Bi da.Detection of Tomato ring spot virus by real-time fluorescent RT-PCR one step assay[J].Acta Phytopathologica Sinica,2003,33(4):338-341. |
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| Authors: | ZHU Jian yu ZHU Shui fang LIAO Xiao lan GAO Bi da |
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| Institution: | 1 Department of Plant Protection, Hunan Agricultural University, Changsha 410128, China;2 Plant Quarantine Lab of State Exit-Entry Inspection and Quarantine Bureau, Beijing 100029, China |
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| Abstract: | A pair of primers and a TaqMan probe based on the conserved nucleotide sequence of RNA1 RNA replicase gene of several ToRSV strains were designed and synthesized. A novel real time fluorescent RT PCR was established to detect ToRSV which was transmited by seed.This technology uses a TaqMan probe to monitor in real time amplification of the target gene.The simultaneous probe detection and PCR amplification obviates any post PCR manipulations and has many advantages,including eliminating ethidium bromide staining and gel running, reducing contamination risks.This reliable, sensitive, quick and easy handling detection method is suitable for diseased diagnosis and quarantine detection. |
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| Keywords: | Tomato ring spot virus seed borne disease real time fluorescent RT PCR detection |
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