畜牧兽医学报2018，Vol.49Issue(5)：996-1004,9.DOI:10.11843/j.issn.0366-6964.2018.05.014
二重荧光RT-LAMP方法鉴别检测口蹄疫病毒和水泡性口炎病毒
Development of a Duplex Fluorescence-based Reverse Transcript Loop-mediated Isothermal Amplification Assay for Detection of Foot-and-mouth Disease Virus and Vesicular Stomatitis Virus
摘要
Abstract
Foot-and-mouth disease and vesicular stomatitis are serious cute disease common in cattle,and prevalence in large parts of the word.The aim of this study was to establish a duplex fluorescence-based reverse transcript loop-mediated isothermal amplification assay for detection of foot-and-mouth disease virus (FMDV) and vesicular stomatitis virus (VSV).Two sets of specific primers were designed according to the conserved sequences of FMDV 3D gene and VSV N gene,and the inner primers were synthesized with two different fluorophores at its 5'ends.A duplex fluorescence-based reverse transcript loop-mediated isothermal amplification assay was developed and used for detection of FMDV and VSV.The results showed that this assay could detect at least 100 copies of premixed plasmid containing the targets gene of FMDV and VSV per tube.The specificity of this assay was high,and be able to detect FMDV and VSV without any crossreactions with other known non-targeted bovine pathogens.Different concentration template of FMDV and VSV could be identified when mixed together without any interference.These findings indicate that this duplex RT-LAMP assay is a simple,rapid,specific,and sensitive method for detection of FMDV and VSV,and can be applied in clinical detection and epidemiological investigation of FMDV and VSV.关键词
口蹄疫病毒/水泡性口炎病毒/二重荧光RT-LAMP/检测Key words
foot-and-mouth disease virus/vesicular stomatitis virus/duplex fluorescence-based RT-LAMP/detection分类
农业科技引用本文复制引用
范晴,邓显文,刘加波,谢芝勋,谢志勤,谢丽基,黄娇玲,张艳芳,曾婷婷,王盛,罗思思..二重荧光RT-LAMP方法鉴别检测口蹄疫病毒和水泡性口炎病毒[J].畜牧兽医学报,2018,49(5):996-1004,9.基金项目
广西科技重大专项(桂科AA17204057) （桂科AA17204057）
