摘要
本研究建立了一种检测传染性造血器官坏死病毒（IHNV）的新型荧光环介导等温扩增法,为实验室及现场检测IHNV提供特异性强、灵敏度高、快速、能实时观察扩增荧光信号的检测技术。针对IHNV全基因组序列设计了一组6条LAMP特异性引物,对4株不同来源的IHNV和8株不同的病毒进行特异性实验和灵敏度实验,结果表明该方法具有高度特异性,检测灵敏度可达3.64×10^-7μg/μL,比常规RT-PCR高1000倍,与荧光RT-PCR方法相当。通过环介导等温扩增技术（LAMP）和荧光检测技术相结合,使其检测时间能缩短至15～20分钟内完成检测,并可杜绝由于开盖加染料而导致的假阳性率偏高问题,在鱼病快速检测上具有重要意义。
A loop-mediated isothermal amplification assay（LAMP）for the detection of infectious haematopoietic necrosis viru（IHNV was established in this study. This novel method could be used in IHNV detection with high specificity,sensitivity and rapidity both in the laboratory and field conditions. A set of 6 LAMP primers were designed based on IHNV genome sequence. 4 IHNVstrains from different sources and 8 other viruses were involved in specificity and sensitivity tests. The results indicated that the specificity was high,and the detection limit was 3.64×10^-7μg/μL,1000 fold higher than the conventional RT-PCR and same as real-time RT-PCR. The combination of LAMP and real-time RT-PCR could decrease the required detection time to 15-20 min and reduce the false positive rate caused by lid opening.
出处
《中国动物检疫》
CAS
2014年第6期68-73,共6页
China Animal Health Inspection
基金
质检公益性行业专项(201210055和201210214)
质检总局科研项目(2012IK032)
