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中国农学通报 ›› 2010, Vol. 26 ›› Issue (23): 18-21.

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

CSFV SIV PCV2 PRV和PRRSV多重PCR检测方法的建立及初步应用

李兆龙 朱小丽 程晓霞 王劭 陈仕龙 林锋强 陈少莺   

  • 收稿日期:2010-07-15 修回日期:2010-08-11 出版日期:2010-12-05 发布日期:2010-12-05
  • 基金资助:

    福建农科院畜牧所所长基金资助

Establishment and application of a multiplex PCR assay for simultaneousdetection of CSFV,SIV,PCV2,PRV and PRRSV

  • Received:2010-07-15 Revised:2010-08-11 Online:2010-12-05 Published:2010-12-05

摘要:

摘 要:参照GenBank中的猪瘟病毒(CSFV)、猪流感病毒(SIV)、猪圆环病毒2型(PCV2)、猪伪狂犬病病毒(PRV)、猪繁殖与呼吸综合病毒(PRRSV)基因序列,设计了5对引物分别用于扩增CSFV E2、SIV、PCV2 ORF2、PRV gB、PRRSV N基因的目的片段。通过对反应条件进行优化,建立了检测CSFV、SIV、 PCV2、 PRV和PRRSV的多重PCR(mPCR)诊断方法。敏感性和特异性结果显示,该mPCR对5种病毒的最低核酸检测量均为106个基因拷贝数(约10 pg)。而对大肠杆菌、葡萄球菌、链球菌、猪细小病毒、鸡传染性喉气管炎病毒、鸡新城疫病毒、猪传染性胃肠炎病毒的多重PCR的扩增结果均为阴性。临床样品的多重PCR检测结果表明,建立的多重PCR方法能够对SIV、CSFV、PCV2、PRV、PRRSV单个或混合感染的临床样品进行快速鉴别诊断。

关键词: 鄂茶, 鄂茶, 品种, 光合特性, 净光合速率

Abstract:

Abstract:According to the gene sequences in GenBank of classical swine fever virus (CSFV), swine influenza virus (SIV), porcine circovirus type 2 (PCV2), porcine pseudorabies virus (PRV), porcine reproductive and respiratory comprehensive virus (PRRSV) gene sequences, designed five pairs of primers were used to amplify CSFV E2, SIV, PCV2 ORF2, PRV gB, PRRSV N gene. Reaction conditions were optimized by the establishment of a detection CSFV, SIV, PCV2, PRV and PRRSV multiple PCR (mPCR) diagnosis. Sensitivity and specificity results showed that the mPCR minimum of 5 of the virus nucleic acid detection volume were 106 gene copy number (about 10 pg). Against E. coli, staphylococcus, streptococcus, porcine parvovirus, infectious laryngotracheitis virus, Newcastle disease virus, porcine transmissible gastroenteritis virus, multiple PCR amplification results were negative. Clinical samples of multiple PCR results showed that the multiplex PCR method capable of SIV, CSFV, PCV2, PRV, PRRSV infection of single or mixed clinical samples for rapid diagnosis. Key words:CSFV;SIV;PCV2;PRV;PRRSV;multiple PCR