摘要
根据GenBank中登录的禽呼肠孤病毒(Aviem reovirus,ARV)S1基因序列设计了特异对应靶序列中的6个基因区段的4条引物,以此建立了一种快速、准确的ARV逆转录环介导等温扩增(RT-LAMP)检测方法,并对此方法的反应条件进行了优化。结果表明,该方法能够特异地扩增ARV,而对其他常见的禽类病毒均无扩增作用,特异性强、重复性好;检测ARV的灵敏度为4ELD50,是RT-PCR方法的10倍;在反应产物中添加SYBR GREENⅠ染料后,可通过肉眼观察有无荧光直接判定结果。
Four specific primers were designed according to the sequences of avian reovirus（ARV） S1 gene available in GenBankTM ,targeting the 6 distinct sequences of S1 gene. A reverse transcriptase loop-mediated isothermal amplication（RT-LAMP） for the rapid detection of ARV was developed by optimizing some of the reaction factors. The results indicated that the sensitivity of the ARV RT-LAMP method was 4 ELDs0, which was 10-fold more sensitive than that of the normal RT-PCR. The method is specific and sensitive. With the addition of SYBR GREEN Ⅰ , the results of RT-LAMP could be detected by naked eyes.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2011年第7期961-964,共4页
Chinese Journal of Veterinary Science
基金
广东省农业产业推进关键技术研究与示范重点资助项目(2009D2009B020201008)
关键词
禽呼肠孤病毒
逆转录环介导等温扩增
条件优化
Avian reovirus （ARV）
reverse transcriptase loop-mediated isothermalamplification （RT-LAMP）
condition optimization
