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本研究以香石竹环斑病毒(Carnation ringspot virus,CRSV)的5个分离物为研究对象,根据CRSV运动蛋白(MP)基因的保守序列设计一对特异性引物和TaqMan荧光探针,建立了检测CRSV的实时荧光RT-PCR(real-timefluorescent RT-PCR)方法。该方法利用TaqMan探针水解产生的荧光信号实时监测目标基因的扩增,实现real-timefluorescent PCR扩增和检测同步进行。结果表明,本研究建立的实时荧光RT-PCR方法具有更快速、灵敏和特异的优点,与普通RT-PCR方法相比其灵敏度提高了100倍,适合于对进境种苗携带的CRSV的快速检测。
In this study, five isolates of Carnation ringspot virus (CRSV) were selected as research objects. A pair of specific primers and TaqMan fluorescent probes were designed according to the conserved sequence of the CRSV gene. CRSV real-time fluorescence RT-PCR (real-timefluorescent RT-PCR) method. The method utilizes the fluorescent signal generated by the hydrolysis of the TaqMan probe to monitor the amplification of the target gene in real time and realizes the amplification and detection of real-time fluorescence amplification in synchronization. The results showed that the real-time fluorescence RT-PCR method established in this study had more rapid, sensitive and specific advantages. Compared with the ordinary RT-PCR method, the sensitivity of the real-time fluorescence RT-PCR method was improved by 100 times, which was suitable for the rapid Detection.