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当前位置: 首页 > 文章 > 菠萝凋萎相关病毒–2实时荧光定量RT-PCR检测方法的建立 园艺学报 2014,41 (6) 1257-1266
Position: Home > Articles > Development of a Real-time Fluorescent Quantitative RT-PCR Method for the Detection of Pineapple mealybug wilt associated virus-2 Acta Horticulturae Sinica 2014,41 (6) 1257-1266

菠萝凋萎相关病毒–2实时荧光定量RT-PCR检测方法的建立

作  者：
胡加谊;罗志文;范鸿雁;李向宏;刘志昕;何凡
单  位：
海南大学环境与植物保护学院;海南省农业科学院热带果树研究所;中国热带农业科学院热带生物技术研究所;农业部海口热带果树科学观测实验站
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
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。
译  名：
Development of a Real-time Fluorescent Quantitative RT-PCR Method for the Detection of Pineapple mealybug wilt associated virus-2
作  者：
HU Jia-yi;LUO Zhi-wen;FAN Hong-yan;LI Xiang-hong;LIU Zhi-xin;HE Fan;College of Environment and Plant Protection,Hainan University;Institute of Tropical Fruit Trees,Hainan Academy of Agricultural Sciences/Haikou Investigation Station of Tropical Fruit Trees,Ministry of Agriculture;Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences;
关键词：
pineapple;;PMWaV-2;;TaqMan probe;;real-time quantitative RT-PCR;;detction
摘  要：
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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