摘要
建立一种快速、敏感、特异的检测H6亚型禽流感病毒(AIV)逆转录环介导等温扩增(RT-LAMP)检测方法。根据H6亚型AIV血凝素(HA)基因序列的保守区8个位点设计了6条特异性LAMP引物,以H6亚型AIV阳性样品RNA为模板进行一步法扩增,对反应条件和反应体系进行优化。结果表明该方法的特异性良好,对其他呼吸道病原体均无扩增反应。该方法灵敏度高,最低可检测到H6亚型AIV RNA为0.01 pg,该方法无需特殊仪器,只需在水浴锅中进行,是一种适于基层的简便、灵敏、快速的H6亚型AIV检测方法。
To develop a rapid sensitive and specific method of a reverse transcription loop-mediated isothermal amplification （RT-LAMP） assay for detection of H6 subtype avian influenza virus （AIV）, six primers specific to the eight site of HA gene were designed according to the sequences of H6 subtype AIV hemagglutinin gene available in GenBank. The viral RNA was amplified with one-step RT-LAMP method, and the reaction conditions were optimized. The results showed that the specificity of the assay was fine without cross amplification of other respiratory tract pathogens, and the detection limit of the RT-LAMP assay was 0.01 pg. The assay is a sensitive, simple, rapid and practical method for detection of H6 subtype AIV in the field.
出处
《微生物学通报》
CAS
CSCD
北大核心
2011年第12期1855-1861,共7页
Microbiology China
基金
国家百千万人才工程人选专项资金项目(No.945200603)
广西科技专项资金项目(No.桂科专项10-3)
广西特聘专家专项经费资助项目(No.2011B020)
