实时荧光RT-PCR方法检测香石竹环斑病毒

Detection of Carnation ringspot virus by real-time fluorescent reverse transcription polymerase chain reaction

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, a pair of specific primers and a TaqMan probe were designed according to the con- served movement protein （MP） gene sequences of Carnation ringspot virus （CRSV）, and a real-time fluores- cent PCR method was applied for detection of CRSV. This method used the fluorescent signal generated from the hydrolysis of TaqMan probe to real-time monitor amplification of the target gene. It realized synchroniza- tion between real-time fluorescent RT-PCR amplification and detection. The results showed that the method was better than others in efficiency and specificity, and 100 times more sensitive than common RT-PCR. This method had potential to be applied in rapid detection of CRSV in incoming seedlings.