猪细小病毒7型Taqman实时荧光PCR检测方法的建立与应用
Development and Application of Taqman Real-time PCR Method for Detection of Porcine Parvovirus 7
  
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中文摘要:
      猪细小病毒7型(Porcine parvovirus 7,PPV-7)是近年新发现的一种细小病毒亚型。为建立PPV-7快速准确的检测方法,以PPV-7病毒全基因组为模板,设计合成1对引物和1条Taqman探针,建立了PPV-7 Taqman实时荧光定量PCR检测方法。本方法的标准曲线分析显示,其常数为0.999 2,敏感性可以达到46个病毒拷贝/μL,批内和批间重复性试验的变异系数均小于2%。用猪圆环病毒2型、3型,猪细小病毒1型,猪伪狂犬病病毒等病原进行特异性试验,证实本方法不能从这些病毒中检测出特异性条带,表明本研究建立的PPV-7实时荧光PCR方法具有特异性强、敏感性高、重复性好的特点。用临床样品对该方法进行了应用性评价,从96份样品中检出52份PPV-7阳性样品,证实了本方法在生产实际应用的可行性。
英文摘要:
      Porcine parvovirus 7(PPV-7)is a subtype of parvovirus newly discovered in recent years. In order to establish a rapid and accurate detection method for PPV-7,a pair of primers and a Taqman probe were designed and synthesized using the whole genome of PPV-7(KU563733)as a template,and a real-time quantitative PCR method for the detection of PPV-7 was established. The standard curve analysis showed its constant was 0.999 2 and the minimum detection limit was 46 copies/μL. For the intra-assay and inter-assay reproducibility tests,all the coefficients of variation were less than 2%. Using the developed method to conduct specificity tests,specific bands couldn't be detected from porcine circovirus type 2 and 3,porcine parvovirus type 1 and porcine pseudorabies virus. These results indicated that the PPV-7 real-time fluorescent PCR method had strong specificity,high sensitivity and reliable reproducibility. Its application was also evaluated by usage of clinical samples. Among 96 samples,52 of them were detected positive for PPV-7,which further confirmed the wide feasibility of this method.
作者单位
张志,张丽丽,刘爽,单虎,李晓成,王树双  
中文关键词:  猪细小病毒7型  实时荧光定量聚合酶链式反应  Taqman探针
英文关键词:porcine parvovirus 7  real time PCR  Taqman probe
基金项目:国家科技基础性工作专项(2012FY111000)
DOI:10.3969/j.issn.1005-944X.2018.09.023
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