首页|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

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

扫码查看
原文链接
  • 国家科技期刊平台
  • NETL
  • NSTL
  • 万方数据
  • 维普
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.
A Quantitative PCR Method for Detecting Phytophthora parasitica var. nicotianae in Tobacco Planting Soil Based on LNA Sensitization
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.

Tobacco planting soilPhytophthora parasitica var. nicotianaeLocked nucleic acid (LNA)Fluorescence quantitative PCRMolecular detection

刘芳、宋纪真、范艺宽、牟文君、奚家勤、胡利伟

展开 >

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

中国烟草总公司河南省公司,郑州市政七街7号 450008

植烟土壤 烟草黑胫病菌 锁核苷酸(LNA) 荧光定量PCR 分子检测

中国烟草总公司河南省公司科技项目

122013DS0320

2015

10.16135/j.issn1002-0861.20151203

烟草科技
中国烟草总公司郑州烟草研究院

烟草科技

CSTPCDCSCD北大核心EI
影响因子：1.049
ISSN：1002-0861
年,卷(期)：2015.48(12)
  • 4
  • 6