猪圆环病毒3型实时荧光定量PCR方法的建立和应用  

Development and Application of A Real-time PCR for Detection of Porcine Circovirus 3

作　　者：郝占武 张志[2] 张美晶[2] 单虎[1] 李晓成[2] HAO Zhan-wu;ZHANG Zhi;ZHANG Mei-jing;SHAN Hu;LI Xiao-cheng(Qingdao Agricultural University,Qingdao,Shandong,266109,China;China Animal Health and Epidemiology Center,Qingdao,Shandong,266032,China)

机构地区：[1]青岛农业大学,山东青岛266109 [2]中国动物卫生与流行病学中心,山东青岛266032

出　　处：《动物医学进展》2018年第8期27-31,共5页Progress In Veterinary Medicine

基　　金：科技基础性工作专项项目(2012FY111000)

摘　　要：为实现猪圆环病毒3型(PCV3)的快速检测,根据GenBank中公布的PCV3全基因组序列设计并合成1对引物,建立了基于SYBR GreenⅠ的实时荧光定量PCR(real-time PCR)检测方法。以前期检测的PCV3阳性样品制备的质粒DNA为模板,绘制了该real-time PCR的标准曲线和熔解曲线分析,并进行了敏感性、特异性和重复性试验。结果表明,该方法具有良好的特异性和较高的敏感性,质粒DNA稀释至10-8后(浓度为5×102拷贝/μL)仍然可以检出,可重复性试验中各组的变异系数均小于3%。用23份临床样品进行了应用性检测,结果从中检测出了5份阳性样品,进一步证实该方法的可行性。A SYBR GreenⅠreal-time PCR for the detection of porcine circovirus 3(PCV3)was developed using a pair of specific primers according to some genomes of PCV3 in the GenBank.A previous plasmid was constructed as a positive template to generate the standard and melting curves.The results showed that this assay had good specificity and high sensitivity for the plasmid even with dilution of 10-8(5×10 2 copies/μL).The Coefficient value of each group was also lower than 3%in the repeatability test.The reliability was further verified when 5 positive samples were detected from 23 clinical samples.

关 键 词：猪圆环病毒3型 实时荧光定量聚合酶链反应 建立 

分 类 号：S852.659.2[农业科学—基础兽医学] S854.43[农业科学—兽医学]