Real-time fluorescent quantitative PCR (Polymerase Chain Reaction) method for detecting tobacco common mosaic virus

The invention relates to the field of real-time detection of mosaic viruses, in particular to a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) method for detecting a tobacco mosaic virus, which comprises the following steps of: 1, designing and synthesizing a TMV primer and a probe; 2, extraction of total RNA and RT-PCR (reverse transcription-polymerase chain reaction); 3, identifying a target gene; 4, preparing a plasmid standard substance; and 5, establishing a standard curve. The method disclosed by the invention is quick, simple, short in time consumption, high in sensitivity, convenient in quantitative detection, strong in specificity and reliable in detection result, so that the method disclosed by the invention can be used as an effective means for screening symptomatic and hidden TMV in tobacco seedlings. 本发明涉及花叶病毒实时检测领域，具体涉及一种用于检测烟草普通花叶病毒的实时荧光定量PCR方法，包括以下步骤：步骤一：TMV引物及探针的设计与合成；步骤二：总RNA的提取与RT-PCR；步骤三：目的基因鉴定；步骤四：质粒标准品的制备；步骤五：标准曲线的建立。本发明的方法快速简单，耗时短，灵敏度高，定量检测方便，特异性强，检测结果可靠，使得本发明的方法可作为烟