多重RT-PCR一步法技术同时检测猪瘟病毒和蓝耳病病毒方法的建立以及初步应用  

Establishment and primary application of multiplex polymerase chainreaction for detection of Classical Swine Fever and Porcine Reproductive and Respiratory Syndrome Virus

作　　者：孔繁德[1] 王荣[2] 陈琼[3] 吴德峰[2] 徐淑菲[1] 

机构地区：[1]厦门出入境检验检疫局,福建厦门361012 [2]福建农林大学动物科学学院,福州350002 [3]厦门市农产品质量安全检验测试中心,福建厦门361009

出　　处：《福建畜牧兽医》2008年第S1期61-65,共5页Fujian Journal of Animal Husbandry and Veterinary medicine

基　　金：厦门出入境检验检疫局科技项目(XK09-2003)

摘　　要：猪瘟病毒和蓝耳病病毒均能导致猪繁殖障碍,对养猪生产影响很大。根据猪瘟病毒(CSFV)和猪蓝耳病(PRRS)的基因保守序列设计2对针对这2种病毒的特异引物,并建立多重RT-PCR方法,分别对其最佳反应条件、特异性及敏感性进行测定。结果表明,能同时扩增得到2条与试验设计相符的167bp(CSFV)和320bp(PRRS)特异性条带,同时具有较好的特异性;敏感性检测结果表明,临床阳性的样品提取的核酸稀释1000倍后仍能检测出CSFV和PRRSV。本方法的建立对于这2种病毒病的早期快速检测具有十分重要的意义。A multiplex polymerase chain reaction (M-PCR) was established and optimized to detect swine fever virus (CSFV) and porcine reproductive and respiratory syndrome virus (PRRSV) simultaneously.Two sets of specific primers were designed according to the conservative sequence of CSFV and PRRSV in the GenBank.It was showed that all sampies which contained CSFV or PRRSV could be amplified by the multiplex PCR using these two sets primers.Two specific bands of CSFV 167 bp and PRRSV 320 bp were detected using agarose gel electrophoresis in this multiplex PCR and accorded with designed result.It could detect 1 000 diluted nu- cleic acid of CSFV or PRRSV from clinic samples. The method provides a fast and reliable way of identifying CSFV or PRRSV. The method could prove very useful for laboratories working with early rapid identification of CSFV or PRRSV.

关 键 词：猪瘟病毒 蓝耳病 多重PCR 

分 类 号：S858.28[农业科学—临床兽医学]