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Position: Home > Articles > Development of Rapid Assay for H7N9 Influenza Virus by Loop-Mediated Isothermal Amplification Method China Animal Health Inspection 2013,30 (7) 72-77

H7N9禽流感病毒等温扩增快速检测方法的建立

作  者：
董志珍;郑文杰;王乃福;赵祥平;吴冬雪;王玉玲;马晶;贾润清;王建华
单  位：
天津出入境检验检疫局动植物与食品检测中心;中国疾病预防控制中心病毒病预防控制所传染病预防控制国家重点实验室
关键词：
H7N9;禽流感病毒;等温扩增
摘  要：
目的建立H7N9禽流感病毒环介导等温扩增(LAMP)快速检测方法。方法从GISAID数据库中获得H7N9禽流感病毒血凝素(HA)和神经氨酸酶(NA)基因序列,应用DNAStar软件MegAlign程序分析其序列,利用PrimerExplorer V4软件在序列保守区域设计LAMP引物,即外引物、内引物,同时以所克隆的阳性质粒为模板,建立H7N9禽流感病毒等温扩增快速检测方法。结果LAMP检测方法对H7N9禽流感病毒的灵敏度达到10个拷贝,所用引物对于H1、H3、H5亚型禽流感病毒和新城疫病毒无非特异性扩增,表现出良好特异性。结论建立的H7N9禽流感病毒等温扩增快速检测方法灵敏度高、特异性强,为快速检测禽流感病毒提供了新方法。
译  名：
Development of Rapid Assay for H7N9 Influenza Virus by Loop-Mediated Isothermal Amplification Method
作  者：
Dong Zhizhen 1 , Zheng Wenjie 1 , Wang Naifu 1 , Zhao Xiangping 1 , Wu Dongxue 1 , Wang yuling 1 , Ma Jing 2 , Jia Runqing 2 , Wang Jianhua 1 , Zhang Xiaoguang 2 , (1.Tianjin Entry-Exit Inspection and Quarantine Bureau,Tianjin 300456; 2.State Key Laboratory for Infectous Disease Prevention and Control, National Institute for Vial Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052)
关键词：
H7N9; AIV; LAMP
摘  要：
Objective To set up a loop-mediated isothermal amplification(LAMP)assay for quick detection of H7N9 avian influenza virus. Methods The HA gene and NA gene of avian influenza virus subtype H7N9 were downloaded from GISAID database and analyzed using MegAlign tools of software DNAStar.The outer primers and inner primers of LAMP were then designed according to their conserva- tive region using software PrimerExplorer V4. The fragment of subtype H7N9 sequence was also ampli- fied and cloned to serve as the positive standard and its LAMP reaction condition and reaction system were optimized in this experiment.Results The results showed that the LAMP assay had a detection limit of 10 DNA copies. The primers used in this assay showed the specificity for H7N9 avian influenza A virus and did not amplify H1、H3、H5 subtype AIV or Newcastle disease virus.Conclusion It is indicated that the LAMP assay is a simple, rapid, accurate, sensitive and specific method for detecting H7N9 AIV. This assay offers a new method and idea for detecting avian influenza virus.

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