畜牧兽医学报2012，Vol.43Issue(4)：659-663,5.
禽新型黄病毒RT-LAMP检测方法的建立
Establishment of Rapid Reverse Transcription Loop Mediated Isothermal Amplification for Detection of Avian Novel Flavivirus
摘要
Abstract
A reverse transcription loop mediated isothermal amplification (RT-LAMP) assay was developed for detection of avian novel flavivirus virus. According to the sequences of avian novel flavivirus virus E protein gene, six primers specific to the eight sites of novel flavivirus virus gene were designed, and the reaction conditions were optimized. The results showed that the assay had no cross reaction with NDRV, GPV, MDRV, NDV, ILTV and FPV. The assay was performed in water bath within 45 minutes and the amplification result was visualized by adding SYBR Green I or inspecting the white sediment. The detection limit of RT-LAMP assay was 1 pg, which was 100 fold higher than that of one step RT-PCR. The detection in clinical samples was 100% compliance with RT-PCR. These results suggested that the newly developed RT-LAMP assay is a simple and specific method for rapid detection of avian novel flavivirus virus in field conditions.关键词
禽新型黄病毒/RT- LAMP/检测方法Key words
avian novel flavivirus/ loop-mediated isothermal amplification/ diagnosis method分类
农业科技引用本文复制引用
李兆龙,陈仕龙,林锋强,王劭,程晓霞,朱小丽,陈少莺..禽新型黄病毒RT-LAMP检测方法的建立[J].畜牧兽医学报,2012,43(4):659-663,5.基金项目
福建省农科院创新团队项目(STIF-Y02) （STIF-Y02）
福建省公益类科研院所专项(2011R1025-2) （2011R1025-2）
