烟草靶斑病菌LFD-RAA快速检测方法的建立
S43; 为快速有效检测烟草靶斑病菌,以烟草靶斑病菌beta-tubulin基因序列保守区域为靶标位点,设计重组酶介导扩增技术(Recombinase aided amplification,RAA)引物,结合侧流层析试纸条(Lateral flow dipstick,LFD),建立烟草靶斑病菌快速检测体系并进行优化.结果表明,LFD-RAA检测体系在39 ℃恒温条件下反应30 min即可完成RAA扩增,并具有较高的特异性,其检测极限值为100 fg/μL,与PCR检测灵敏度相当.对检测体系优化确定检测扩增的最优时间为20～35 min、扩增温度为31～39 ℃.LFD-RAA检测体系可应用于...
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| Published in | 烟草科技 Vol. 57; no. 5; pp. 41 - 47 |
|---|---|
| Main Authors | , , , , , , |
| Format | Journal Article |
| Language | Chinese |
| Published |
湖南省烟草公司郴州市公司,湖南省郴州市北湖区燕泉街道燕泉北路61号 423000%湖南省烟草科学研究所,长沙市芙蓉南路368号 410004%湖南省烟草公司永州市公司,湖南省永州市冷水滩区珍珠北路69号 425000%湖南农业大学植物保护学院,长沙市芙蓉区农大路1号 410128
01.05.2024
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1002-0861 |
| DOI | 10.16135/j.issn1002-0861.2023.0408 |
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| Abstract | S43; 为快速有效检测烟草靶斑病菌,以烟草靶斑病菌beta-tubulin基因序列保守区域为靶标位点,设计重组酶介导扩增技术(Recombinase aided amplification,RAA)引物,结合侧流层析试纸条(Lateral flow dipstick,LFD),建立烟草靶斑病菌快速检测体系并进行优化.结果表明,LFD-RAA检测体系在39 ℃恒温条件下反应30 min即可完成RAA扩增,并具有较高的特异性,其检测极限值为100 fg/μL,与PCR检测灵敏度相当.对检测体系优化确定检测扩增的最优时间为20～35 min、扩增温度为31～39 ℃.LFD-RAA检测体系可应用于烟草上烟草靶斑病菌的快速检测. |
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| AbstractList | S43; 为快速有效检测烟草靶斑病菌,以烟草靶斑病菌beta-tubulin基因序列保守区域为靶标位点,设计重组酶介导扩增技术(Recombinase aided amplification,RAA)引物,结合侧流层析试纸条(Lateral flow dipstick,LFD),建立烟草靶斑病菌快速检测体系并进行优化.结果表明,LFD-RAA检测体系在39 ℃恒温条件下反应30 min即可完成RAA扩增,并具有较高的特异性,其检测极限值为100 fg/μL,与PCR检测灵敏度相当.对检测体系优化确定检测扩增的最优时间为20～35 min、扩增温度为31～39 ℃.LFD-RAA检测体系可应用于烟草上烟草靶斑病菌的快速检测. |
| Abstract_FL | To rapidly and effectively detect the tobacco target spot pathogen,the conserved region of the beta-tubulin gene sequence of the pathogen was targeted and the recombinase aided amplification(RAA)primers were designed.Lateral flow chromatographic strips were used to establish and further optimize a rapid detection system.The results showed that the developed RAA amplification could be completed within 30 minutes at a constant temperature of 39 ℃ and had high specificity with a detection limit of 100 fg/μL.Its sensitivity was comparable to that of conventional PCR detections.The optimal amplification time and temperature of the optimized detection system were 20-35 min and 31-39 ℃.This LFD-RAA detection system could be applied for rapid detection of the pathogen of tobacco target spot on tobacco plants. |
| Author | 肖艳松 李思军 钟杰 吴文信 周向平 刘天波 周陆苏 |
| AuthorAffiliation | 湖南省烟草公司郴州市公司,湖南省郴州市北湖区燕泉街道燕泉北路61号 423000%湖南省烟草科学研究所,长沙市芙蓉南路368号 410004%湖南省烟草公司永州市公司,湖南省永州市冷水滩区珍珠北路69号 425000%湖南农业大学植物保护学院,长沙市芙蓉区农大路1号 410128 |
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| Author_FL | ZHONG Jie LI Sijun ZHOU Lusu WU Wenxin ZHOU Xiangping XIAO Yansong LIU Tianbo |
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| Keywords | 烟草靶斑病 重组酶介导扩增技术 Detection 检测 Tobacco target spot Recombinase aided amplification |
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| Snippet | S43; 为快速有效检测烟草靶斑病菌,以烟草靶斑病菌beta-tubulin基因序列保守区域为靶标位点,设计重组酶介导扩增技术(Recombinase aided amplification,RAA)引物,结合侧流层析试纸条(Lateral flow... |
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| Title | 烟草靶斑病菌LFD-RAA快速检测方法的建立 |
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