套式RT-PCR检测牛呼吸道合胞体病毒的研究  

Detection of bovine respiratory syncytial virus by a reverse transcriptase-nested-polymerase chain reaction in bovine clinical samples

作　　者：史鸿飞[1] 朱远茂[1] 高欲燃[1,2] 任宪刚[1] 冯军科[1] 于作[1] 薛飞[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/大动物传染病研究室,黑龙江哈尔滨150001 [2]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2010年第3期238-240,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省"十一五"科技攻关项目(GA06B202-3)

摘　　要：为了检测牛呼吸道合胞体病毒(BRSV),根据已发表的融合蛋白(F)基因序列,设计了套式RT-PCR引物,初步建立了BRSV的套式RT-PCR检测方法。对138份牛鼻腔棉拭子进行了检测,结果检测到了BRSV阳性样品54份,总的阳性检出率为39.1%。对大部分BRSV阳性样品的F基因扩增产物进行了序列测定与分析。用该套式RT-PCR对牛传染性鼻气管炎病毒、牛副流感病毒3型、牛病毒性腹泻病毒等进行了检测,结果无交叉反应,表明该检测方法具有良好的特异性。本研究首次用套式RT-PCR技术证实了我国部分省的牛群中存在BRSV感染。A nested RT-PCR for bovine respiratory syncytial virus （BRSV） detection with two pairs of primers was developed based on the fusion （F） gene according to the published nucleotide sequences of BRSV. A total of 138 clinical nasal swab samples were detected using the nested RT-PCR for BRSV,and 54 of 138 samples were shown to be positive （39.1 %）. The amplified fragments of the F gene from the most samples were sequenced and analyzed. No cross-reactions were found in samples of infectious bovine rhinotracheitis virus,bovine influenza virus type 3,and bovine viral diarrhea virus. These results indicated that the nested RT-PCR was specific for BRSV detection. This is the first detection of BRSV by the nested RT-PCR in clinical samples from some provinces in China.

关 键 词：牛呼吸道合胞体病毒 套式RT-PCR 牛鼻腔拭子 

分 类 号：S852.65[农业科学—基础兽医学]