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依据甘薯羽状斑驳病毒(Sweet potato feathery mottle virus,SPFMV)外壳蛋白(CP)基因的保守区设计引物和TaqMan探针,经过对反应体系和反应条件的优化,建立了特异、灵敏、高效的SPFMV实时荧光定量PCR检测方法。试验结果表明:该方法能检测到目的病毒,标准曲线的斜率和相关系数分别为-3.307和0.998,扩增效率为100.6%。最低可检测到约5.46个拷贝的阳性质粒,灵敏度比常规PCR高2个数量级。建立的实时荧光定量PCR方法可用于田间样品的检测,为SPFMV的早期预警和流行学研究提供了技术手段。
According to the conserved region of CP gene of sweet potato feathery mottle virus (SPFMV), primers and TaqMan probes were designed. After optimization of the reaction system and reaction conditions, a specific, sensitive and efficient SPFMV Real-time fluorescence quantitative PCR detection method. The experimental results show that the method can detect the target virus. The slope and the correlation coefficient of the standard curve are -3.307 and 0.998 respectively, and the amplification efficiency is 100.6%. A minimum of 5.46 copies of positive plasmids could be detected with 2 orders of magnitude higher sensitivity than conventional PCR. The established real-time fluorescence quantitative PCR method can be used for the field sample detection, which provides a technical measure for the early warning and epidemiological study of SPFMV.