摘要
根据H7N9亚型禽流感病毒(AIV)HA基因和NA基因的保守序列,分别设计特异性引物和不同荧光基团标记的TaqMan探针。通过优化反应条件,建立了H7N9亚型AIV的双重实时荧光定量RTPCR检测方法。结果显示,该法检测H7N9亚型AIV的下限为102copies/μL,批内重复和批间重复变异系数均小于3%。本研究建立的H7N9亚型AIV双重实时荧光定量RT-PCR方法,具有快速、特异和敏感的优点,可为H7N9亚型AIV的有效防控提供技术支撑。
The specific primers and TaqMan probes were designed according to the conserved sequences of the hemagglutinin（HA）genes of H7 subtype AIV and the neuramidinase（NA）genes of N9 subtype AIV,respectively.The reaction conditions were optimized to develop a duplex real-time RT-PCR assay for rapid detection of H7N9 AIV.It was shown that the specificity of this assay was high,the amplification curves were displayed for the detection of H7N9 AIV,H7 subtype AIV and N9 subtype AIV,and no positive results were observed when nucleic acid from other subtypes AIV and avian pathogens.The detection limit of this assay was 100 copies of H7N9 AIV,and the coefficients of variation were both less than 3% for the intra-assay and inter-assay.This duplex assay is a rapid,specific and sensitive method for the detection of H7N9 AIV,and provides a technical support to prevent and control H7N9AIV.
出处
《动物医学进展》
CSCD
北大核心
2013年第12期1-5,共5页
Progress In Veterinary Medicine
基金
广西特聘专家专项(2011B020)
广西科技攻关重大专项(桂科重1222003-2-4)
广西科技攻关(1010014-5)
