禽流感病毒和新城疫病毒二联RT-PCR检测方法的建立  

Establishment of AIV and NDV by Multiplex RT-PCR

作　　者：于洋[1] 李敬双[1] 徐长顺[1] 吴健[1] 周铁忠[1] 于金玲[1] 孙党章[1] 史丽华[1] 

机构地区：[1]锦州医学院畜牧兽医学院,辽宁锦州121001

出　　处：《动物医学进展》2006年第4期84-86,共3页Progress In Veterinary Medicine

基　　金：辽宁省教育厅高等学校科学研究项目(2004D259)

摘　　要：参照国内外已发表的禽流感病毒(AIV)和新城疫病毒(NDV)的基因序列及其相关的RT-PCR检测方法,根据禽流感病毒M蛋白基因和新城疫病毒NP基因各设计一套特异性通用引物,扩增目的带分别为600bp和340 bp。通过对相关病毒检测,建立了AIV和NDV通用型二联RT-PCR检测方法。该方法具有快速、敏感、特异等优点,可为AIV和NDV的检测、流行病学调查及疫苗使用等奠定基础。According to the published gene sequence of Avian influenza virus （AIV） and Newcastle disease virus （NDV） and the detecting methods of RT-PCR reported, matrix（M） gene of AIV and nucleoprotein （NP）gene of NDV were considered as the target gene sequences of multiple RT-PCR. A set of universe primers were designed for detecting AIV and NDV, respectively in the conserved M gene of AIV and NP gene of NDV. The products of RT-PCR is 600 bp and 340 bp respectively. The optimal condition of multiple RT-PCR for detection of AIV and NDV by RT-PCR was determined through orthogonal assay. The specificity of the multiple primers were examined by RT-PCR using template extracted from other avian virus. The sensitivity of RT-PCR were also determined by a seral dilution of RNA from AIV and NDV. This study have paved the way for testing AIV and NDV, defining pathogenic potential, using vaccines against AIV and NDV and so on.

关 键 词：禽流感病毒 新城疫病毒 反转录聚 合酶链式反应 

分 类 号：S852.659.5[农业科学—基础兽医学]