摘要
为建立一种H6亚型禽流感病毒(AIV)的套式RT-PCR检测方法,根据GenBank中H6亚型AIV HA基因序列,设计了4条特异性引物,优化反应条件,并对所建立的方法进行特异性和敏感性的检验,用该法对154份活禽市场样品进行检测。结果表明,所建立的套式RT-PCR方法对其他常见禽病病原体无扩增;对H6亚型AIV的最小检测限为1×102拷贝/μL,灵敏度比常规RT-PCR方法高100倍;154份样品的检测结果与病毒分离一致。所建立的H6亚型AIV套式RT-PCR检测方法具有特异性强和灵敏度高的特点。
This study aimed to establish a specific and sensitive nested RT-PCR for detection of H6 subtypes avian influenza virus（AIV）.Four specific primers were designed according to HA gene sequences of H6 subtype AIV in Genbank,and the reaction conditions were optimized.To further evaluate the reliability,the specificity and sensi-tivity of assay were evaluated,and 154 clinical samples were detected by nested RT-PCR.The results showed that the specificity of the assay was high without amplification of the other viruses.The detection limit of nested RT-PCR was 1×102 copies/μL,which was 100-fold higher than that of the routine PCR.In the detection of 154 clini-cal samples,the results of this assay were accorded with the viral isolation completely.In conclusion,this assay is a specific and sensitive method,and also provides an effective method to detect for the H6 subtype AIV.
出处
《动物医学进展》
CSCD
北大核心
2014年第11期10-13,共4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)
广西畜禽疫苗新技术重点实验室项目(12-071-28-A-01)
