| 用地高辛标记的核酸探针检测猪伪狂犬病毒野毒感染的研究 |
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| 引用本文： | 刘志杰,任慧英,温建新,刘文华,邹玲,韩先杰,魏笑笑. 用地高辛标记的核酸探针检测猪伪狂犬病毒野毒感染的研究[J]. 畜牧兽医学报, 2008, 39(11) |
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| 作者姓名： | 刘志杰 任慧英 温建新 刘文华 邹玲 韩先杰 魏笑笑 |
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| 作者单位： | 青岛农业大学动物科技学院,青岛,266109 |
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| 基金项目： | 山东省青岛市科技攻关项目 |
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| 摘 要： | 利用PCR技术从带有伪狂犬病毒（PRV）gE基因的重组质粒pMD18-T-gE中扩增回收约304bp大小的片段，并制备出地高辛标记的gE基因核酸探针。特异性检测结果表明，该探针能与重组质粒DNA发生特异性杂交，而与对照的PRVBartha-k61株疫苗毒DNA、猪细小病毒（PPV）DNA、猪圆环病毒（PCV）DNA、猪繁殖与呼吸综合征病毒（PRRsV）cDNA、猪瘟病毒（CSFV）cDNA的杂交反应均为阴性；敏感性检测结果表明，该探针对PRV野毒的最低检出量为4pg。应用该探针对11份繁殖障碍病料进行了杂交检测，共检出4份阳性病料，该结果与PCR检测结果一致，表明该核酸探针可用于猪伪狂犬病野毒感染的临床诊断。
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| 关 键 词： | 伪狂犬病毒 gE基因 地高辛标记探针 斑点杂交 野毒株 |
Detection of Wild-type Porcine Pseudorabies Virus by Digoxigenin-labeled DNA Probe |
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| LIU Zhi-jie,REN Hui-ying,WEN Jian-xin,LIU Wen-hua,ZOU Ling,HAN Xian-jie,WEI Xiao-xiao. Detection of Wild-type Porcine Pseudorabies Virus by Digoxigenin-labeled DNA Probe[J]. Chinese Journal of Animal and Veterinary Sciences, 2008, 39(11) |
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| Authors: | LIU Zhi-jie REN Hui-ying WEN Jian-xin LIU Wen-hua ZOU Ling HAN Xian-jie WEI Xiao-xiao |
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| Affiliation: | LIU Zhi-jie,REN Hui-ying,WEN Jian-xin,LIU Wen-hua,ZOU Ling,HAN Xian-jie,WEI Xiao-xiao(College of Animal Science , Veterinary Medicine,Qingdao Agricultural University,Qingdao 266109,China) |
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| Abstract: | A 304 bp fragment of gE gene of PRV was amplified from recombinant plasmid pMD18-T-gE and labeled with digoxigenin as DNA Probe.Dot blot hybridization was developed for detection of PRV wild strains.The recombinant plasmid DNA,PRV Bartha-k61DNA,PPV DNA,PCV DNA,PRRSV cDNA and CSFV cDNA were tested by this method.The results showed that positive signal appeared only in recombinant plasmid,while PRV Bartha-k61 DNA,PPV DNA,PCV DNA,PRRSV cDNA and CSFV cDNA were all negative.As little as 4 pg of known positive ta... |
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| Keywords: | pseudorabies virus gE gene digoxigenin-labeled probe dot blot hybridization wild-type strain |
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