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 TaqMan probe based on specific primers of conserved nucleotide sequence of PMWaV-2 coat protein gene. The results showed that the method was higldy sensitive to positive sample, but had no fluorescence signal to health sample and blank control. The sensitivity of RTqPCR was about 100 times higher than general PCR. Reproducibihty test revealed that the coefficients of variation between intra-and inter-assay were beth within 1.85%, indicating it was a quantitative detection method for PMWaV-2 with simple operation, strong specificity, high sensitivity, and reliable reproducibility.
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 TaqMan probe based on specific primers of conserved nucleotide sequence of PMWaV-2 coat protein gene. The results showed that the method was higldy sensitive to positive sample, but had no fluorescence signal to health sample and blank control. The sensitivity of RTqPCR was about 100 times higher than general PCR. Reproducibihty test revealed that the coefficients of variation between intra-and inter-assay were beth within 1.85%, indicating it was a quantitative detection method for PMWaV-2 with simple operation, strong specificity, high sensitivity, and reliable reproducibility.
基金
Supported by Applied Research and Industrialization Projects of Key Science and Technology Plan of Hainan Province(ZDXM20130031)
Special Fund for Agro-scientific Research in the Public Interest(201203021)
Scientific Operation Fund of Hainan Province(QCY[2013]131)
Natural Science Foundation of Hainan Province(QK[2013]32)
Key Research and Development Project of Hainan Province(ZDYF2016035)
Special Program for Agricultural Science and Technology Innovation of Hainan Academy of Agricultural Sciences(CXZX201410)
