摘要
根据猪圆环病毒2型(PCV2)BF的序列设计引物,扩增472bp的ORF2部分基因片段,并克隆到pGEM-T载体中,构建含PCV2-ORF2部分基因片段的重组质粒。利用XbaⅠ限制性内切酶消化后获得共用同1对引物但缺失196bp的竞争重组子。通过重组质粒与竞争重组子间的竞争定量PCR方法(qc-PCR),建立猪PCV2-ORF2的标准竞争曲线和直线回归方程,并对PCV2试验感染猪血清中PCV2核酸的动态变化进行了研究。
An 472 bp fragment of porcine circovirus type 2 （PCV2） was amplified from the total DNA of the PCV2 BF strain by PCR method, and one recombinant （primitive plasmid） was obtained by ligation of the 472 bp fragment and pGEM-T vector. The other recombinant （competitive plasmid） contained 276 bp fragment of PCV2- ORF2 which can be amplified by the same pair of primer for the 472 bp fragment,was constructed by Xba I digestion with the premititive plasmid. By the quantitative competitive PCR between the logarithm of ratios of amplified copies of primitive plasmid （which will be replaced by cDNA of PCV2 DNA） to the amplified copies of series 2-fold dilution competitive plasmid copies in comparison with the molecules of series 2 fold dilution competitive plasmid, the linear regression equation of the standard competitive curve for PCV2 DNA detection was verified, meanwhile, the kinetics of viral load in sera of PCV2 experimentally infected pigs were detected.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2008年第10期1115-1117,共3页
Chinese Journal of Veterinary Science
基金
教育部优秀跨世纪优秀人才计划资助项目(2003)
关键词
猪圆环病毒2型
竞争定量PCR
病毒含量
porcine circovirus virus type 2 （PCV2）
quantitative competitive PCR （qc-PCR）
viral load
