摘要
根据GenBank中的羊传染性脓疱病毒(ORFV)B2L基因序列,设计合成1对引物,通过对PCR条件的优化,敏感性、特异性试验,成功建立了ORFV的PCR检测方法。敏感性与特异性结果显示,最低核酸检测量为1.2 fg/μL,对口蹄疫病毒、山羊痘病毒、丝状支原体山羊亚种的扩增结果均为阴性。同时根据GenBank中的ORFV的CBP基因的序列,设计合成1对特异性引物,以ORFV贵州分离株作为模板,成功克隆出ORFV的CBP基因。同源性分析表明,核苷酸与GenBank中ORFV的CBP基因的核苷酸序列相似性99.4%～88.8%。本研究为ORFV感染的流行病学调查和CBP基因功能研究奠定基础。
According to the gene sequences of B2L gene of Orf virus（ORFV） in GenBank,one pair of specific primers was designed for amplifying the gene fragment of ORFV.The PCR method for detection of ORFV was established by the optimization of reaction conditions.The sensitivity of the established PCR was high and the detection limit was 1.2fg/μL of ORFV DNA.No band was amplified from foot and mouth disease virus（FMDV）,goatpox virus（GPV） and Mycoplasma mycoides spp.capri（Mmc）.The CBP gene was amplified from ORFV Guizhou strain,and sequence analysis showed 99.4%~88.8% of identities with the CBP gene sequences in GenBank.This study provides a basis for molecular epidemiology of ORFV infection and function of CBP gene.
出处
《畜牧与兽医》
北大核心
2013年第7期30-34,共5页
Animal Husbandry & Veterinary Medicine
基金
贵州省农业科技攻关项目[黔科合NY字(2009)3089号]
贵州省畜禽健康养殖技术创新能力建设项目[黔科合院所创能(2010)4004]
中央补助地方科技基础条件专项基金项目[黔科条中补地(2010)4001]
黔西南州种草养羊产业发展省州科技合作专项项目2012[3]号
