摘要
根据GenBank中登录的H7亚型禽流感病毒(H7AIV)血凝素基因序列,设计了一套特异识别HA基因序列中6个不同区段的环介导恒等温扩增(LAMP)引物,并以此套引物建立了一种基于反转录环介导等温核酸扩增(RT-LAMP)技术的H7亚型禽流感病毒诊断方法。结果显示,该方法对H7AIV RNA的最小检测限为0.1pg,灵敏度高于普通一步法RT-PCR 100倍;全部反应可在1.5h内完成;在反应体系中添加Fluorescent Detection Reagent染料后,可通过肉眼观察有无荧光直接判定结果。灵敏性及特异性试验证明,该方法灵敏度高、特异性好,能够作为H7亚型禽流感病毒的快速诊断方法。
A series of specific primer were designed according to the sequences of H7 subtype avian influenza virus（H7 AIV） hemagglutinin（HA） gene available in GenBank. A reverse transcriptase loop-mediated isothermal amplification（RT-LAMP） for the rapid detection of H7 AIV was developed using specific primers. In result,the detection limit of the developed RT-LAMP method was 100-fold higher than that of the routine one-step RT-PCR. The amplification could be finished within 1.5 h with good specificity and high sensitivity. These results indicated that the developed RT-LAMP method could facilitate the rapid diagnosis of avian influenza subtype H7.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2013年第12期1224-1230,共7页
Chinese Veterinary Science
基金
中央级公益性科研院所基本科研业务费专项:禽流感RT-LAMP诊断技术的研究(ZGKJ201005)
关键词
H7亚型禽流感病毒
反转录环介导恒等温扩增
快速检测
H7 subtype avian influenza virus
reverse transcriptase loop-mediated isothermal amplifica- tion（RT-LAMP）
rapid detection
