摘要
针对猪脑心炎病毒(Encephalomyocarditis virus,EMCV)3D基因保守序列的6个特异性部位设计了2对引物,利用Bst DNA聚合酶,63℃恒温保持50min即可完成反转录与扩增反应,产物中加入SYBR GreenⅠ染料,在紫外光下可直接观察判定扩增结果,试验成功建立了猪EMCV的RT-LAMP检测方法。结果表明:该方法灵敏度比RT-PCR高100倍,具有良好的重复性和稳定性,而且对其他病毒均无扩增结果,可作为猪EMCV的快速诊断方法。
A reverse transcription-loop-mediated isothermal amplification（RT-LAMP） assay for encephalomyocarditis virus （EMCV） was established with two pairs of primers based on identified 6 positions in 3D gene conserved sequence. The process of assay was conducted with Bst DNA polymerase at a constant temperature of 63 ℃ per 50 min, and the products could be visualized under the UV light with SYBR Green I dye. The result shows that：the sensi- tivity of RT-LAMP assay was 100 times higher than RT-PCR,with preferable reproducibility and it was not able to detect the other virus. Thus,we could conclude that this established LAMP assay has a potential application to the rapid detection of EMCV.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2012年第1期44-47,共4页
Chinese Journal of Veterinary Science
关键词
猪脑心肌炎病毒
3D基因
环介导逆转录等温扩增
porcine encephalomyocarditis virus
3D gene
reverse transcription-loop-mediated isothermal amplification（RT-LAMP）
