猪圆环病毒3型半巢式PCR方法的建立和应用
Establishment and Application of Half Nested-PCR for Porcine Circovirus 3
  
查看全文  查看/发表评论  下载PDF阅读器
中文摘要:
      为建立快速检测猪圆环病毒3型(PCV-3)的方法,以GenBank上登陆的我国PCV-3全基因组为参考毒株,设计3条引物,建立了 PCV-3半巢式PCR诊断方法。本方法可以扩增出大小为249 bp的特异性条带,与预期大小一致,而猪圆环病毒1型和2型、猪伪狂犬病病毒、猪细小病毒等对照样品均未扩增出特异性条带。将PCV-3的阳性DNA稀释后作为模板,发现半巢式PCR的检测极限可以达到5×10-3 μg/mL。对临床80份样品进行检测,发现半巢式PCR的阳性检出率可以达到27.5%(22/80),而常规PCR的检出率仅为6.25%(5/80)。检测结果表明,本文建立的半巢式PCR方法较为准确、特异和敏感,适用于PCV-3的快速检测和流行病学调查。
英文摘要:
      In order to establish a method for rapid detection of Porcine circovirus type 3(PCV-3),a new half nested-PCR method was constructed to detect the virus of Porcine circovirus 3(PCV-3)on the base of three primers designed by the conference strain genome from the GenBank. The special band of 249 bp was amplified as the same size as expected. The specific bands were not amplified in the control samples,including PCV-1,PCV-2,PRV and PPV. After series dilution of positive DNA of PCV-3,the available limit of dilution DNA could reach 5×10-3 μg/mL to amplify the special band. Meanwhile,application of 80 clinical samples also showed that the positive rate of half nested-PCR could reach 27.5%(22/80),while the detection rate of conventional PCR was only 6.25%(5/80). The results indicated that the half nested-PCR in this research was accurate,sensitive and special,and was suitable for rapid detection and epidemiology investigation of PCV-3.
作者单位
张志,郝占武,杨旭兵,张美晶,吴发兴,李晓成  
中文关键词:  猪圆环病毒3型  半巢式-聚合酶链式反应  快速检测  流行病学调查
英文关键词:Porcine circovirus 3  half nested-PCR  quick detection  epidemiological survey
基金项目:科技部科技基础性工作专项(2012FY111000)
DOI:10.3969/j.issn.1005-944X.2018.01.023
关闭

“中国兽医发布”微信公众号
以下是在线客服