期刊论文

Detection of Burkholderiapseudomalleiin soil by real-time fluorescence loop-mediated isothermal amplification

中文

中国人兽共患病学报

1002-2694

2019

35

3

216-222

10.3969/j.issn.1002-2694.2019.00.004

国家重点研发计划(No.2017YFC1601503) 中国载人航天预先研究项目(No.010101) 湖南教育厅重点课题(No.15A165) 南华大学大学生研究性学习与创新性实验计划(No.20164890101),南华大学自然科学基金项目资助(No.2018KYY266) 衡阳市重点实验室项目(No.2018KJ110)联合资助.

本校为第一机构

目的结合荧光染料EvaGreen与环介导等温扩增技术(Loop-mediated isothermal amplification,LAMP)探索快速检测类鼻疽伯克霍尔德菌的可行性。方法根据类鼻疽伯克霍尔德菌filc基因片段设计了LAMP引物,利用荧光染料EvaGreen建立实时荧光环介导等温扩增技术(real-time fluorescence loop-mediated isothermal amplification,RealAmp),优化反应条件,扩增产物经电泳鉴定。通过类鼻疽伯克霍尔德菌和其他致病菌验证特异性,比较RealAmp与普通LAMP技术的敏感度,并测定人工污染土壤模拟样本的检出限。结果恒温63℃条件下,20~60 min即可完成RealAmp检测;利用该方法检测类鼻疽伯克霍尔德菌为阳性,其余致病菌如...

The purpose of this study is using fluorescent dye EvaGreen and loop-mediated isothermal amplification technology to explore the feasibility of the rapid detection of Burkholderia pseudomallei.The specific LAMP primers designed were based on the filc gene of Burkholderia pseudomallei,and the best reaction conditions with EvaGreen inside for the real-time fluorescence loop-mediated isothermal amplification were optimized,and the products of amplification were detected by electrophoresis.Comparison of the sensitivity between RealAmp and general L...