中国畜牧兽医

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检测猪伪狂犬病病毒和猪细小病毒的二重PCR方法的建立

韩勇   

  1. 山东省日照市动物疫病预防控制中心,山东日照 276800
  • 收稿日期:2012-03-21 出版日期:2012-09-20 发布日期:2012-09-18
  • 作者简介:韩勇(1980-),男,山东人,硕士生,研究方向:动物传染病学与分子生物学。

Establishment of Double PCR Method for Detecting Porcine Pseudorabies Virus and Porcine Parvovirus

HAN Yong   

  1. Animal Epidemic Prevention and Control Center of Rizhao in Shandong Province,Rizhao 276800,China
  • Received:2012-03-21 Online:2012-09-20 Published:2012-09-18

摘要: 作者建立了一种同时检测猪细小病毒(PPV)和猪伪狂犬病病毒(PRV)的二重PCR方法。根据GenBank上发表的PPV和PRV基因序列,针对各自保守区各设1对特异性引物,用这2对引物对同一样品中的PPV和PRV进行检测,结果同时扩增出942 bp(PPV)和485 bp(PRV)2条特异性片段。特异性试验表明,对其他几种病原的PCR扩增结果均为阴性。敏感性试验结果表明,该二重PCR能检出PPV 和PRV最低浓度分别为22、11.7 pg/L。该方法的建立对临床上进行这2种疾病的鉴别诊断和混合感染的检测都具有重要意义。

关键词: 二重PCR; 猪细小病毒; 猪伪狂犬病病毒

Abstract: A double PCR was optimized to simultaneousely detect two pathogens of PPV and PRV. Two sets of primers were designed according to the conservative gene sequences of PPV and PRV. It was proved that the DNA samples which contained PPV and PRV could be amplified by the double PCR using the two sets of primers and it would yield two specific fragments of 942(PPV) and 485 bp(PRV).But no specific fragments were amplified from other pathogenic viruses and bacterium. The sensitive determination result indicated that as little as PPV 22 pg/L and PRV 11.7 pg/L could be detected. This method could differentiate PRV,PPV and mixed infection from the reproductive failure.

Key words: double PCR; PPV; PRV

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