实时荧光PCR检测马铃薯白线虫技术研究

马铃薯胞囊线虫是马铃薯上最重要的有害生物之一,也是我国特别关注的重要植物检疫性线虫.针对马铃薯白线虫ITS序列,我们设计了引物和TaqMan探针,使用15种马铃薯胞囊线虫群体和4种其它胞囊线虫样品进行验证,可高度灵敏地检测单个马铃薯白线虫的胞囊或幼虫,最高检测灵敏度达到10fs;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探针特异性.该检测方法可自动化检测马铃薯白线虫并进行定量,适合进行标准化的常规检测.

Real time PCR of Globodera pallida with TaqMan probe

Potato cyst nematodes are one of the most important plant pests infested potatoes, also are plant quar-antine nematodes concerned specially by Chinese Government. To carry out rapid and reliable species detection method of Globodera pallida, we designed primers and TaqMan probe depended on sequence of ITS. The primers and probe were evaluated on 15 samples of PCN populations and 4 other cyst nematodes. The PCR assay for spe-cies detection is highly sensitive,which could detect the template concentration as low as 10 fg. This method can identify and quantify single air - dried cyst and juvenile of G. pallida, and also allows the detection of species mixtures in a sample. Furthermore, the realtime PCR assay with TaqMan provides a rapid means for simultaneous detection and quantification of PCN and is therefore particularly useful for routine diagnoses.