Detection of Pineapple mealybug wilt associated virus-3 by Real-Time Fluorescent Quantitative RT-PCR

【Objective】The objective of this paper was to establish the method of Real-Time Fluorescent Quantitative PCR(q PCR) for quantitative detection of pineapple mealybug wilt associated virus- 3(PMWa V3).【Method】Specific Taq Man probe and primers were designed and synthesized according to the conserved sequence of coat protein(CP) gene of Pineapple mealybug wilt associated virus-3(PMWa V-3),and the standard curve was established after optimizing the amplification condition of q PCR.【Result】The results showed that the method was specific for the detection of PMWa V-3. Meanwhile,the research revealed that the sensitivity of the present method was about 10 times higher compared to general conventional RT-PCR and the variation coefficients of intra-assay and inter-assay were less than 1.73% respectively.【Conclusion】The study suggested that q PCR is an easy,fast and reliable method for the quantitative detection of PMWa V-3.