A SYBR Green Ⅰ Real-time Fluorescence Quantitative PCR Detection Method for the Mulberry MosaicDwarf associated Virus

【Objective】The purpose of this study was to establish a real-time fluorescence quantitative PCR detection method for the Mulberry Mosaic Dwarf associated Virus (MMDaV).【Method】As target gene，the conservative coat protein gene of MMDaV was applied to design the specific primers，construct the positive plasmids of MMDaV and establish the standard curve of the positive plasmids.The sensitivity and specificity of the particular primer of MMDaV was detected.The real-time fluorescence quantitative PCR detection method of MMDaV was applied to detect MMDaV of diseased mulberry leaf from six provinces of Guangdong，Guangxi，Hainan，Chongqing，Shaanxi，and Jiangxi.【Results】 The standard curve constructed has excellent amplification efficiency (97.88%).The specific primer can accurately detect the MMDaV.The detection method has the lowest plasmid detection concentration of 8 copies/μL，the sensitivity of which was 24 times higher than that of conventional PCR.The method has good detection results for diseased mulberry leaf samples from six provinces，and the Cq value of detection is between 9.69 and 26.17.【Conclusion】The established real-time fluorescence quantitative PCR detection method for MMDaV，with high efficiency，specificity and sensitivity，can be applied to detect MMDaV in host plants quantitatively.