【摘要】 非洲猪瘟病毒p54蛋白在病毒感染与增殖过程中起重要作用。根据p54基因的核苷酸序列,设计并合成引物和用6-carboxy-fluorescein(6-FAM)和tetramethyl-6-carboxyrhodamine(TAMRA)荧光素标记TaqMan双水解探针,以构建的含p54基因的pET-p54重组质粒作为阳性模板,建立了ASFV检测的荧光定量PCR方法(已申请专利,专利号200910067620.6)。结果表明,所建立的方法与含p54基因的模板质粒数呈现很好的相关性(R2=0.991),可检测出低于15个拷贝/μL的样品,灵敏度和特异性高,可作为出入境检验检疫部门对非洲猪瘟病毒的快速检测方法。更多还原

【Abstract】 African swine fever virus(ASFV) protein p54 plays an important role in the viral infection and proliferation.Using the recombinant plasmid pET-p54 as template and TaqMan probe,a real-time fluorescence quantitative PCR assay to detect p54 gene of the ASFV was developed with 6-carboxy-fluorescein(6-FAM) and tetramethyl-6-carboxyrhodamine(TAMRA) reporter dyes(patent NO: 200910067620.6).The standard curve was created by detecting the quantitative recombinant plasmid pET-p54.The result showed that the developed method was specific and sensitive,the sensitivity can be as low as 15 gene copies,and the correlation coefficient of the standard curve is up to 0.991.All these data suggested that this method might be useful in daily entry-exit inspection of the ASFV.更多还原