黄瓜细菌性角斑病菌交叉引物恒温扩增检测技术

Detection of Pseudomonas syringae pv. lachrymans by cross priming amplification method

黄瓜细菌性角斑病是我国黄瓜生产上的重要病害之一,其病原菌为Pseudomonas syringae pv.lachrymans。根据该病原菌甘油醛-3-磷酸脱氢基因保守序列设计引物和探针,建立了交叉引物恒温扩增和核酸试纸条检测技术。菌体DNA检测灵敏度可达0.55 ng,纯菌直接检测灵敏度基本可达到单个细菌。所测试的5株黄瓜细菌性角斑病菌和染病黄瓜叶片均为阳性,其他13株对照菌株均为阴性。该方法灵敏度高,且操作简单,对设备要求低等,适合基层实验室应用。

Angular leafspot of cucumber is one of the most important diseases on cucumber caused by Pseudomonas syringae pv. lachrymans,which is destructive and widespread. To develop a cross-priming and nucleic acid strip method for detection of P. syringae pv. lachrymans,primers and probes set was designed based on glyceraldehyde-3-phosphate dehydrogenase 1（gap 1） gene. The result showed that all the five P. syringae pv. lachrymans strains and infected cucumber leaves were positive,whereas thirteen other relative pathogens strains were negative. The limitation on DNA detection is 0.55 ng,and it can detect individual bacteria in culturing. Thus,this method has advantages of high sensitivity,good specificity,easy operation and low requirements of expensive equipments,is suitable for basic level laboratory.