Establishment of a Real-time Fluorescent Quantita- tive RT-PCR Method for Pineapple mealybug wilt as. sociated virus-2

• 作者: Fan Hongyan Guo Lijun He Fan He Shu Hu Fuchu Hu Jiayi Li Xianghong Luo Zhiwen Zhang Zhiyi
• 来源期刊: Plant Diseases and Pests(植物病虫害研究：英文版)
• 学科: 农学
• 关键词: RT-PCR方法 实时定量PCR 菠萝 荧光定量 粉蚧 枯萎 病毒 特异性引物
• 年，卷（期）: 2016,（1）
• 页码范围: 22-26
• 页数: 5页
• 分类号: S858.31

摘要

Pineapple mealybug wilt associated virus-2 (PMWaV-2) is an important pathogen causing Mealybug wilt of pineapple (MWP). We established a realtime quantitative RT-PCR( RT-qPCR) method with Taq Man probe based on specific primers of conserved nucleotide sequence of PMWaV-2 coat protein gene. The results showed that the method was highly sensitive to positive sample,but had no fluorescence signal to health sample and blank control. The sensitivity of RT-qPCR was about 100 times higher than general PCR. Reproducibility test revealed that the coefficients of variation between intra-and inter-assay were both within 1.85%,indicating it was a quantitative detection method for PMWaV-2 with simple operation,strong specificity,high sensitivity,and reliable reproducibility.