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文章编号: 16726987(2019)03002904; DOI: 10.16351/j.16726987.2019.03.005
王熊, 周鑫昱, 史文 强, 刘彩艳, 李荣贵*, 石超*
(青岛大学 生命科学学院,山东 青岛 266071)
摘要: 利用新型的等温扩增技术实现了松材线虫的快速检测。研究中通过改进核酸提取方法,只需95 ℃处理5 min就能完成线虫核酸释放。通过在反应开始前加入核酸扩增显色染料,实现了扩增结果的肉眼可视化判定。实验结果表明;整个等温扩增反应过程只需50 min,可以检测单条线虫,反应只需恒温进行。因此,该方法为松材线虫的快速检测提供了一种简单、有效的工具。
关键词: 松材线虫; 等温核酸扩增; 快速检测; 可视化
中图分类号: Q 819文献标志码: A
引用格式: 王熊, 周鑫昱, 史文 强, 等. 一种松材线虫快速检测的新技术\[J\]. 青岛科技大学学报(自然科学版), 2019, 40(3): 2932.
WANG Xiong, ZHOU Xinyu, SHI Wenqiang, et al. A new technique for the rapid detection of nucleic acid in pine wood nematode\[J\]. Journal of Qingdao University of Science and Technology(Natural Science Edition), 2019, 40(3): 2932.
A New Technique for the Rapid Detection of Nucleic Acid in Pine Wood Nematode
WANG Xiong, ZHOU Xinyu, SHI Wenqiang, LIU Caiyan, LI Ronggui, SHI Chao
(Department of Biology, Qingdao University, Qingdao 266071, China)
Abstract: A novel isothermal amplification technique was used to rapidly detect pine wood nematodes. In the study, an improved nucleic acid extraction method was used. The nucleic acid of the nematode was released at 95 ℃ for 5 min. The nucleic acid amplification chromogenic dye was added before the start of the reaction, and the amplification result could be directly determined by visual observation. The experimental results showed that the whole isothermal amplification reaction process could be finished in 50 min, and single nematodes could be detected at constant temperature. This method provides a convenient, effective tool for the rapid detection of pine wood nematodes.
Key words: pine wood nematode; isothermal amplification; rapid detection; visualization
收稿日期: 20180516
基金项目: 山东省重点研发计划项目(2016GNC110024);国家自然科学基金项目(21675094).
作者简介: 王熊(1992—),男,硕士研究生.*通信联系人.