摘要
【目的】建立一种可以定量检测猪圆环病毒3型(PCV3)的荧光定量PCR检测方法。【方法】设计与验证1对PCV3特异性引物,以系列稀释后的PCV3全基因组质粒为模板进行SYBR Green I荧光定量PCR扩增。【结果】该方法不能检出PCV1与PCV2等常见猪DNA病毒,精确灵敏度至少达到100 copies,所绘制标准曲线回归方程的决定系数为0.999,扩增效率为83.6%,对53份临床样品的检测阳性率高于常规PCR。【结论】建立1种具有良好特异性、敏感性与重复性的定量检测PCV3的荧光定量PCR方法。
【Objective】The present study was to establish a fluorescent quantitative PCR(FQ-PCR)assay for quantitative detection of porcine circovirus type 3(PCV3).【Methods】A pair of primers specific to PCV3 was designed and validated,SYBR Green I real-time PCR was performed using a series of diluted complete genome plasmid of PCV3 as templates.【Results】Using this generated method,epidemic swine DNA viruses such as PCV1 and PCV2 could not be detected,the sensitivity reached at least 100 copies,the determinant coefficient for the regression equation of the standard curve was 0.999,the amplification efficiency was 83.6%,the positive rate of 53 clinical samples was higher than that of conventional PCR.【Conclusion】A FQ-PCR assay for quantitative detection of PCV3 with good specificity and sensitivity was established.
作者
王阳
胡胜云
石玉佩
申贞彦
孙彤
周双海
WANG Yang;HU Shengyun;SHI Yupei;SHEN Zhenyan;SUN Tong;ZHOU Shuanghai(College of Animal Science and Technology,Beijing University of Agriculture,Beijing 102206,China)
出处
《北京农学院学报》
2020年第2期29-33,共5页
Journal of Beijing University of Agriculture
基金
北京市科技计划课题(D171100002117002)。
作者简介
第一作者:王阳,硕士生,主要从事兽医传染病学研究,Tel:010-80799397,E-mail:840569091@qq.com;第一作者:胡胜云;通信作者:周双海,副教授,主要从事兽医传染病学研究,Tel:010-80799397,E-mail:shhaizhou@sina.com。
