| 魏梅生，田茜，赵文军* , 李桂芬，孔君.番茄细菌性叶斑病菌RPA检测技术研究[J].植物保护,2016,42(1):150-153. |
| 番茄细菌性叶斑病菌RPA检测技术研究 |
| Rapid detection of Pseudomonas syringae pv. tomato by RPA method |
| 投稿时间：2014-12-14 修订日期：2015-01-03 |
| DOI： |
| 中文关键词: 番茄细菌性叶斑病菌 重组酶聚合酶扩增 RPA 等温扩增 检测 |
| 英文关键词:Pseudomonas syringae pv. tomato recombinase polymerase amplification RPA isothermal amplification detection |
| 基金项目:质检公益性行业科研专项(201310071); “十二五”国家科技支撑计划项目(2012BAK11B02) |
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| 中文摘要: |
| 番茄细菌性叶斑病菌(Pseudomonas syringae pv. tomato, Pst)是我国进境植物检疫性有害生物, 可随种子进行远距离传播。快速简便的检测对于防止该病害的扩散传播具有重要意义。依据番茄细菌性叶斑病菌的hrpZPst 基因序列, 设计并筛选出特异性扩增引物, 建立了番茄细菌性叶斑病菌的重组酶聚合酶等温扩增(RPA)检测方法。该方法可从10种不同的植物病原细菌中特异性地检测到3个参试番茄细菌性叶斑病菌株。对病菌DNA的检测灵敏度为75 fg/μL, 与PCR凝胶电泳相当。该方法扩增核酸时间短、效率高、对设备的要求低, 适合于基层简易实验室的快速检测。本文为该病原菌的检测提供了新方法。 |
| 英文摘要: |
| Bacterial speck of tomato caused by Pseudomonas syringae pv. tomato (Pst) is a serious disease in tomato and a threat to tomato production, which could be transmitted by infected seeds. Pst is considered a quarantine organism and subject to phytosanitary regulations in China. Rapid and convenient detection method is important for the disease control. A recombinase polymerase amplification (RPA) assay was developed based on the hrpZPst gene sequence. Three isolates of Pst can be specifically detected and other nine bacterial strains got negative result. The detection limit of RPA for bacterial genome DNA is about 75 fg/μL, similar to PCR agarose gel electrophoresis. The method is rapid, sensitive, and suitable for use in primary level laboratory. |
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