[Objective] The paper aimed to establish a real-time fluorescent quantitative PCR( qPCR) detection method for Pineapple mealybug wilt associated virus-3( PMWa V3). [Method] Specific TaqMan probe and primers were designed and synthesized according to the conserved sequence of coat protein( CP) gene of PMWa V-3,and the standard curve was established after optimizing the amplification condition of qPCR. [Result]The results showed that the method was specific for the detection of PMWa V-3,and the sensitivity of the present method was about 10 times higher compared to general RT-PCR; the variation coefficients of intraassay and inter-assay were less than 1. 73,respectively. [Conclusion] The qPCR is an easy,fast and reliable method for quantitative detection of PMWa V-3. [Objective] The paper aimed to establish a real-time fluorescent quantitative PCR (qPCR) detection method for Pineapple mealybug wilt associated virus-3 (PMWa V3). [Method] Specific TaqMan probe and primers were designed and synthesized according to the conserved sequence of coat protein (CP) gene of PMWa V-3, and the standard curve was established after optimizing the amplification condition of qPCR. [Result] The results showed that the method was specific for the detection of PMWa V-3, and the sensitivity of the present method was about 10 times higher than to general RT-PCR; the variation coefficients of intraassay and inter-assay were less than 1. 73, respectively. [Conclusion] The qPCR is an easy, fast and reliable method for quantitative detection of PMWa V-3.