实时荧光技术在鸭坦布苏病毒RT-LAMP检测方法中的应用
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- 英文论文题名：Application of real-time fluorescent technique for the detection of duck Tembusu virus by reverse transcription loop-mediated isothermal amplification
- 论文作者：董嘉文 ; 孙敏华 ; 李林林 ; 袁建丰 ; 胡奇林 ; 张建峰
- 英文论文作者：Dong Jiawen ; Sun Minhua ; Li Linlin ; Yuan Jianfeng ; Hu Qilin ; Zhang Jianfeng ; Institute of Animal Health ; Guangdong Academy of Agriculture Sciences ; Guangdong Open Laboratory of Veterinary Public Health ; Guangdong Provincial Key Laboratory of Livestock Disease Prevention
- 年：2016
- 作者机构：广东省农业科学院动物卫生研究所广东省兽医公共卫生公共实验室广东省畜禽疫病防治研究重点实验室;
- 论文关键词：鸭坦布苏病毒 ; 逆转录环介导等温扩增 ; 实时荧光技术
- 英文论文关键词：Duck Tembusu virus ; reverse transcription loop-mediated isothermal amplification ; real-time fluorescent technique
- 会议召开时间：2016-06-14
- 会议录名称：第25届广东省科技进步活动月畜牧兽医学术与科技创新发展大会论文集
- 语种：中文
- 分类号：S852.65
- 学会代码：GDKL
- 会议名称：第25届广东省科技进步活动月畜牧兽医学术与科技创新发展大会
- 会议地点：中国广东广州
- 主办单位：广东省畜牧兽医学会
- 学会名称：广东省科学技术协会科技交流部
- 页数：7
- 文件大小：1136k
- 原文格式：O
- 会议级别：地方
摘要
2010年4月以来,由鸭坦布苏病毒(DTMUV)引起的蛋鸭产蛋急剧下降给我国的养鸭业造成了巨大的经济损失。本研究针对DTMUV E基因保守区域设计一套LAMP引物,利用实时荧光技术建立了DTMUV的实时RT-LAMP病原检测方法。该方法检测RNA的最低检测极限可达到7.8 copies/μL,其灵敏度是普通RT-PCR的100倍,且只能特异性扩增鸭坦布苏病毒RNA,对其他常见鸭源病毒核酸均无特异性扩增。该方法简单快速、特异性强和灵敏度高,判定结果只需要观察扩增曲线,适用于DTMUV早期感染的诊断、分子流行病学调查和疫情监测。
Duck Tembusu virus(DTMUV) has caused huge losses to the poultry industry in China since April of 2010.A loop-mediated isothermal amplification(LAMP) assay based on real-time fluorescent technique was developed for the rapid detection of DTMUV using a set of specific primers according to the conserved region of DTMUV E gene.The sensitivity of real-time RT-LAMP was 100 times more sensitive than the conventional RT-PCR,with a detection limit of 7.8 copies/μL.The specificity of the real-time RT-LAMP was confirmed using RNAs and DNAs extracted from related viruses which cause duck infections.This established method was specific and sensitive,can quickly detect pathogen by observing the amplification curve.As a practical molecular diagnostic method,it can be used for rapid detection of DTMUV early infection and epidemiological investigation.
Duck Tembusu virus(DTMUV) has caused huge losses to the poultry industry in China since April of 2010.A loop-mediated isothermal amplification(LAMP) assay based on real-time fluorescent technique was developed for the rapid detection of DTMUV using a set of specific primers according to the conserved region of DTMUV E gene.The sensitivity of real-time RT-LAMP was 100 times more sensitive than the conventional RT-PCR,with a detection limit of 7.8 copies/μL.The specificity of the real-time RT-LAMP was confirmed using RNAs and DNAs extracted from related viruses which cause duck infections.This established method was specific and sensitive,can quickly detect pathogen by observing the amplification curve.As a practical molecular diagnostic method,it can be used for rapid detection of DTMUV early infection and epidemiological investigation.
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