H7亚型禽流感病毒实时荧光定量RT-PCR检测方法的建立  

Development of Real-time RT-PCR for Avian Influenza Virus Subtype H7

作　　者：卢体康[1,2] 秦智锋[1,3] 陈兵[1] 阮周曦[1] 陶虹[1] 吕建强[1] 花群义[1,3] 孙洁[1] 曾少灵[1] 曹琛福[1] 张彩虹[1] 

机构地区：[1]深圳出入境检验检疫局,广东深圳518045 [2]深圳市外来有害生物质检测技术研发重点实验室,广东深圳518045 [3]深圳市检验检疫科学研究院,广东深圳518001

出　　处：《动物医学进展》2012年第3期9-13,共5页Progress In Veterinary Medicine

基　　金：国家质检总局科研项目(2012IK0034)

摘　　要：H7亚型禽流感病毒致病性强、危害性大,是进出口家禽及禽类产品的主要检疫对象。为了加强对H7亚型禽流感病毒的检测,建立新的快速检测方法。通过对GenBank已报道的H7亚型禽流感病毒的HA基因进行序列分析比较,设计了两套分别针对H7N2亚型和H7Nx亚型的特异性引物和用FAM标记的Taq Man MGB核酸探针,建立了H7亚型禽流感病毒实时荧光定量RT-PCR方法(Real-time RT-PCR)。该方法特异性好,不存在假阴性和假阳性的现象;敏感性高,对禽流感病毒H7亚型标准HI抗原检测的敏感性达到10-5,能够满足口岸禽流感病毒快速、准确、有效的检疫需求。The subtype H7 of avian influenza virus is the major quarantine disease in the avian and avian products in international trade because it is a high exotic pathogenic infectious disease with great impact on poultry production.In order to rapidly test H7 subtype in laboratory,a Taq Man real-time reverse transcription-PCR for detecting H7 subtype influenza virus directed at the HA gene was developed specifically with two sets of primers.One set of primers and probe were special to H7N2 subtypes and the other was to H7Nx.The sensitivit of real-time RT-PCR assay was 10-5 dilution of H7 HI standard antigen.The specificity of assay developed had no cross reaction with others.These methods potentially allowed for more rapid,sensitive,and specific detectiing avian products in trade.

关 键 词：禽流感病毒 H7亚型 实时荧光定量RT-PCR 检测 

分 类 号：S855.3[农业科学—临床兽医学]