| 菠萝凋萎相关病毒–2实时荧光定量RT-PCR检测方法的建立 |
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| 引用本文： | 胡加谊,罗志文,范鸿雁,李向宏,刘志昕,何,凡.菠萝凋萎相关病毒–2实时荧光定量RT-PCR检测方法的建立[J].园艺学报,2014,41(6):1257-1266. |
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| 作者姓名： | 胡加谊 罗志文 范鸿雁 李向宏 刘志昕 何 凡 |
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| 作者单位： | （1海南大学环境与植物保护学院，海口 570228；2海南省农业科学院热带果树研究所，农业部海口热带果树科学观测实验站，海口 571100；3中国热带农业科学院热带生物技术研究所，海口 571101） |
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| 摘 要： | 菠萝凋萎相关病毒（Pineapple mealybug wilt associated virus-2，PMWaV-2）是引起的菠萝凋萎病（Mealybug wilt of pineapple，MWP）的重要病原。本研究中根据PMWaV-2外壳蛋白基因序列设计特异性引物和探针，建立基于TaqMan探针的实时荧光定量RT-PCR检测方法。该方法能高灵敏检测出阳性样品，对阴性样品及空白对照均无荧光反应；灵敏度比普通PCR高100倍；重复性试验表明批内和批间变异系数均在1.85%以内，表明本方法是一种操作简便、特异性强、灵敏度高、重复性较好的PMWaV-2定量检测方法。
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| 关 键 词： | 菠萝 菠萝凋萎病毒–2（PMWaV-2） TaqMan探针 实时荧光定量RT-PCR 检测 |
| 收稿时间： | 2014-01-27 |
Development of a Real-time Fluorescent Quantitative RT-PCR Method for the Detection of Pineapple mealybug wilt associated virus-2 |
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| HU Jia-yi,LUO Zhi-wen,FAN Hong-yan,LI Xiang-hong,LIU Zhi-xin,HE Fan,.Development of a Real-time Fluorescent Quantitative RT-PCR Method for the Detection of Pineapple mealybug wilt associated virus-2[J].Acta Horticulturae Sinica,2014,41(6):1257-1266. |
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| Authors: | HU Jia-yi LUO Zhi-wen FAN Hong-yan LI Xiang-hong LIU Zhi-xin HE Fan |
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| Institution: | （1 College of Environment and Plant Protection，Hainan University，Haikou 570228，China；2 Institute of Tropical Fruit Trees，Hainan Academy of Agricultural Sciences/Haikou Investigation Station of Tropical Fruit Trees，Ministry of Agriculture，Haikou 571100，China；3Institute of Tropical Bioscience and Biotechnology，Chinese Academy of Tropical Agricultural Sciences，Haikou 571101，China） |
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| Abstract: | Pineapple mealybug wilt associated virus-2（PMWaV-2）is an important pathogen that causes Mealybug wilt of pineapple（MWP）. This study established a real-time quantitative RT-PCR method with TaqMan probe based on specific primers of conserved nucleotide sequence of PMWaV-2 coat protein gene. The results showed that the method is highly sensitive to positive sample，but has no fluorescence signal to health sample and water control. The sensitivity of real-time quantitative RT-PCR is about 100 times higher than regular PCR. Three-time repeats revealed that the coefficients of variation between the intra- and inter-assay were both within 1.85%，indicating a simple，specificity，high sensitivity，and reliable reproducibility detection method to PMWaV-2. |
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| Keywords: | pineapple PMWaV-2 TaqMan probe real-time quantitative RT-PCR detction |
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