| 检测猪戊型肝炎病毒的荧光定量PCR方法的建立 |
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| 引用本文： | 张亮权,欧阳昀,刘志刚,曹宗喜,闫明菲,王文华,郭泗虎,张桂红.检测猪戊型肝炎病毒的荧光定量PCR方法的建立[J].中国兽医杂志,2011,47(10). |
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| 作者姓名： | 张亮权 欧阳昀 刘志刚 曹宗喜 闫明菲 王文华 郭泗虎 张桂红 |
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| 作者单位： | 华南农业大学兽医学院,广东广州,510642 |
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| 基金项目： | 国家生猪现代农业产业技术体系，广州市科技支撑计划项目"猪戊型肝炎诊断关键技术的研究" |
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| 摘 要： | 根据GenBank中猪戊型肝炎病毒的ORF2核苷酸序列的保守区域设计合成一对特异性引物,建立了一套SYBRGreen Ⅰ荧光定量PCR检测猪源戊型肝炎病毒(swHEV)的方法,并评价了该方法的灵敏度、稳定性和特异性,同时与常规的RT-nPCR进行对比分析.结果表明,建立标准曲线的相关系数为0.998,斜率为-3.039,Ct值变异系数(CV)在0.17％～1.41％之间,有良好的稳定性.同时在检测猪群常见病中显示出很好的特异性,并且比RT-nPCR更灵敏,适合于swHEV的检测.
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| 关 键 词： | 猪戊型肝炎病毒 荧光定量PCR RT-nPCR |
Establishment of a real-time fluorescent quantitative PCR assay for the detection of swine hepatitis E virus |
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| ZHANG Liang-quan,OUYANG Yun,LIU Zhi-gang,CAO Zong-xi,YAN Ming-fei,WANG Wen-hua,GUO Si-hu,ZHANG Gui-hong.Establishment of a real-time fluorescent quantitative PCR assay for the detection of swine hepatitis E virus[J].Chinese Journal of Veterinary Medicine,2011,47(10). |
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| Authors: | ZHANG Liang-quan OUYANG Yun LIU Zhi-gang CAO Zong-xi YAN Ming-fei WANG Wen-hua GUO Si-hu ZHANG Gui-hong |
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| Institution: | ZHANG Liang-quan,OUYANG Yun,LIU Zhi-gang,CAO Zong-xi,YAN Ming-fei,WANG Wen-hua,GUO Si-hu,ZHANG Gui-hong(College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China) |
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| Abstract: | According to the published sequences of swHEV in GenBank,a pair of primers were designed and synthesized for the conserved open reading frame 2(ORF2) of HEV,and a SYBR Green I fluorescent real-time quantitative PCR assay was developed.The sensitivity,stability and specificity were evaluated,and comparison of the Real-time PCR with nested RT-PCR was performed.According to the experiment,the correlation coefficient(r2)and the slope value of the standard curve was 0.998 and-3.039 respectively.The coefficient o... |
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| Keywords: | swine hepatitis E virus real-time fluorescent quantitative PCR RT-nPCR |
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