应用TaqMan探针进行马铃薯金线虫实时荧光PCR检测技术研究

马铃薯孢囊线虫是全球性的植物检疫性线虫，亦是我国重点关注的有害生物。针对马铃薯金线虫ITS序列，设计了引物和TaqMan 探针，使用15个马铃薯孢囊线虫群体和4个其他孢囊线虫样品进行验证，可高度灵敏地检测单个马铃薯金线虫的孢囊或幼虫，并可进行定量；最高检测灵敏度达到10fg；同时开展了混合样品和未知样品的检测，证明了引物的专化性和TaqMan探针的特异性。该检测方法可自动化检测马铃薯金线虫并进行定量，适合进行标准化的常规检测。

Real time PCR assay of the golden nematode, Globodera rostochiensis with TaqMan probe

Potato cyst nematodes are important plant quarantine pests in the world, and also specially concerned the Chinese government. To carry out rapid and reliable detection of the golden potato cyst nematode Globodera rostochiensis by using real time PCR, we designed primers and probes. The primers and probes were evaluated on 15 samples of PCN populations and 4 other cyst nematodes. The PCR assay for species detection was highly sensitive, which could detect the template concentration as low as 10 fg. This method could identify and quantify air dried and single cyst of the potato cyst nematode, and also allowed detection of species mixtures in a sample. Furthermore, the real time PCR assay with TaqMan provided a rapid means for simultaneous detection and quantification of PCN and is therefore particularly useful for routine diagnoses.