罗非鱼罗湖病毒实时荧光RT-LAMP检测方法的建立

Establishment of Real-time RT-LAMP Detection Method for Tilapia Lake Virus

近年来,罗湖病毒(Tilapia lake virus,TiLV)频繁出现,给罗非鱼养殖业造成巨大损失。本文建立的罗湖病毒实时荧光反转录环介导等温扩增(Reverse transcription loop-mediated isothermal amplification,RT-LAMP)技术,操作简单,检测灵敏度可达30 copies/反应,特异性良好,对其他常见的淡水鱼类病毒核酸无非特异性扩增。31份样本中,阳性3份,阴性28份,检测结果与RT-PCR方法基本一致,两者的符合率为97%。本文建立的TiLV实时荧光RT-LAMP快速诊断新方法,可及时监控、预警我国罗非鱼养殖过程中罗湖病毒(TiLV)的存在,为全面提升我国罗非鱼养殖技术水平和效益提供技术支持。

In recent years,the phenomenon of large-scale death of tilapia caused by tilapia lake virus(TiLV)has resulted in great losses to tilapia aquaculture industry.The real-time fluorescent reverse transcription loop-mediated isothermal amplification technique for TiLV was established in this paper.It found that the real-time fluorescent reverse transcription loop-mediated isothermal amplification technique was characterized by simple operation and sensitivity of 30 copies/reaction,with good specificity of the detection.There has no specific amplification for nucleic acids of other common freshwater fish viruses.The 33 samples actually detected,three positive and 28 negative,were being consistent with those of RT-PCR by 97%.The new method of TiLV real-time RT-LAMP rapid diagnosis established in this paper can timely monitor the existence of TiLV and early warn in tilapia culture,which provides technical support for improving the technical level and success rate of tilapia culture benefits in China.