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禽流感病毒H5和H9亚型RT-PCR检测方法的建立
引用本文:刘燕,钱爱东.禽流感病毒H5和H9亚型RT-PCR检测方法的建立[J].吉林农业大学学报,2004,26(1):93-96.
作者姓名:刘燕  钱爱东
作者单位:吉林农业大学动物科技学院,吉林,长春,130118
基金项目:吉林省科技厅资助项目(20005514),长春市科技局资助项目(20099)
摘    要:根据H5和H9亚型的HA基因,设计合成2对特异性引物,分别建立了RT-PCR检测方法,优化了反应体系及反应条件,该方法敏感性可达10^-3,特异性好,与NFV、IBV与H5与H9相互间无交叉反应。利用所建立的RT-PCR检测了10份活禽及禽产品中存在的H5和H9亚型禽流感病毒,其检测结果与病毒的分离培养一致,比电镜和琼扩的检出率高30%,与其他NDV、IBV的病料无交叉反应,证明该方法具有很好的应用价值。

关 键 词:禽流感病毒  H5亚型  H9亚型  RT-PCR  检测方法  禽流感
文章编号:1000-5684(2004)01-0093-04

Establishing Method of RT-PCR Detecting H5,H9 Subtypes of Avian Influenza Virus
LIU Yan,QIAN Ai-dong.Establishing Method of RT-PCR Detecting H5,H9 Subtypes of Avian Influenza Virus[J].Journal of Jilin Agricultural University,2004,26(1):93-96.
Authors:LIU Yan  QIAN Ai-dong
Institution:LIU Yan,QIAN Ai-dong~
Abstract:Two specific primers were designed according to the HA sequences reported and registered in Genebank of different strains of H_5,H_9 subtypes.The procedure using RT-PCR to detect H_5,H_9 subtypes of AIV was established and the optimal conditions of PCR for each subtype were determined by orthongonal assay. The method is very practable and specific. Its allergy is 10~(-3). It could be used in diagnosis of H_5,H_9 subtypes of AIV.The method was used to detect H_5 and H_9 subtypes of AIV, which hide in poultry and in the products of poultry . The rudimentary application of the RT-PCR has the same results as seperate cultivation of the virus.The positive results acquired were 30%higher than AG and EM.
Keywords:AIV  detection  H_5 subtype  H_9 subtype
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