猪伪狂犬病病毒及猪细小病毒二联PCR检测方法的建立  

Establishment of double PCR for PRV and PPV

作　　者：郁宏伟[1] 杨润德[1] 崔弈杰[1] 秦彤[1] 刘晓慧[1] 

机构地区：[1]河北农业大学动物科技学院,河北保定071001

出　　处：《中国预防兽医学报》2006年第2期216-219,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：河北省科技攻关计划项目(04820403)

摘　　要：根据猪伪狂犬病病毒(PRV)和猪细小病毒(PPV)2种病原体的基因保守序列,分别设计并合成了与PRV的gp50基因序列和PPV的VP2基因互补的两对引物,进行单相PCR,分别扩增出预期的217bp片段及158bp片段;同时,用这两对引物对混合的样品中的PRV和PPV的DNA模板进行二联PCR扩增及二联PCR反应条件的优化,结果得到2条与试验设计相符的217bp(PRV)和158bp(PPV)特异性条带:敏感性检测结果表明,对PRV的检测可以达到10-6.11/20μLTCID50、对PPV的检测可以达到0.008HA单位的核酸模板。本文建立的PCR方法,可以在24h内对样品进行快速检测,可作为PRV、PPV感染和临床样品检测的可靠手段。Two pairs of specific, primers were designed according to the conservative domain of PRV（gp50） and PPV （VP2）.The zone of DNA amplification of single PCR appeared in 217 bp and 158 bp,respectively.Meanwhile,by improving reactive condition,it was showed that two samples which contained PRV and PPV DNA could be amplified by the double PCR using these two sets of primers,yielding two specific bands of 217 bp and 158 bp. As little as 10^-6.11/20 μ L TCID50 and 0.008 HA unit of PPV DNA were detected through sensitization test. The methods could get result in 24h which may be a reliable way in dectecting PRV and PPV.

关 键 词：猪伪狂犬病病毒 猪细小病毒 二联PCR 

分 类 号：S852.655[农业科学—基础兽医学]