| 基于锁核苷酸(LNA)增敏的植烟土壤中烟草黑胫病菌定量PCR检测方法 |
| |
| 引用本文： | 刘芳,宋纪真,范艺宽,牟文君,奚家勤,胡利伟.基于锁核苷酸(LNA)增敏的植烟土壤中烟草黑胫病菌定量PCR检测方法[J].烟草科技,2015,48(12):14-19. |
| |
| 作者姓名： | 刘芳 宋纪真 范艺宽 牟文君 奚家勤 胡利伟 |
| |
| 作者单位： | 1.1. 中国烟草总公司郑州烟草研究院, 郑州高新技术产业开发区枫杨街2号 450001;2.2. 中国烟草总公司河南省公司, 郑州市政七街7号 450008 |
| |
| 基金项目： | 中国烟草总公司河南省公司科技项目“烟草黑胫病防控关键技术研发与应用”(122013DS0320)。 |
| |
| 摘 要： | 为了快速有效地检测植烟土壤中的烟草黑胫病菌的定殖数量,以一种锁核苷酸(LNA)引物为基础,进行了植烟土壤中烟草黑胫病菌数量的定量PCR 检测方法研究。结果表明:①与普通DNA引物相比,LNA 引物提高了PCR 反应的退火温度,减少了引物二聚体和非特异性产物的形成,提高了烟草黑胫病菌分子检测的灵敏度与特异性;②定量分析检测出14 个烟草-大蒜轮作土样中烟草黑胫病菌的定殖数量为2.76×103～5.20×104个/g,且在4～5 h 内完成检测,具有较高的灵敏度和检测效率。
|
| 关 键 词： | 植烟土壤 烟草黑胫病菌 锁核苷酸(LNA) 荧光定量PCR 分子检测 |
| 收稿时间： | 2015-01-19 |
A Quantitative PCR Method for Detecting Phytophthora parasitica var. nicotianae in Tobacco Planting Soil Based on LNA Sensitization |
| |
| LIU Fang,SONG Jizhen,FAN Yikuan,MOU Wenjun,XI Jiaqin,HU Liwei.A Quantitative PCR Method for Detecting Phytophthora parasitica var. nicotianae in Tobacco Planting Soil Based on LNA Sensitization[J].Tobacco Science & Technology,2015,48(12):14-19. |
| |
| Authors: | LIU Fang SONG Jizhen FAN Yikuan MOU Wenjun XI Jiaqin HU Liwei |
| |
| Affiliation: | 1.1. Zhengzhou Tobacco Research Institute of CNTC, Zhengzhou 450001, China;2.2. Henan Province Company of CNTC, Zhengzhou 450008, China |
| |
| Abstract: | In order to efficiently detect the colonization of Phytophthora parasitica var. nicotianae in tobacco planting soil, a quantitative PCR method was researched based on locked nucleic acid (LNA) primer. The results showed that: 1) Comparing with conventional DNA primers, the LNA primer raised PCR reaction annealing temperature, reduced the formation of primer dimers and non-specific products, and improved the sensitivity and specificity of detection. 2)LNA quantitative PCR analysis revealed that the concentration of Phytophthora parasitica var. nicotianae in 14 soil samples of tobacco / garlic rotation fields ranged from 2.76×103 to 5.20×104 per gram. This method is sensitive and efficient, the detection can be completed within 4-5 hours. |
| |
| Keywords: | |
| 本文献已被 CNKI 等数据库收录！ |
| 点击此处可从《烟草科技》浏览原始摘要信息 |
|
点击此处可从《烟草科技》下载全文 |
|
