用分子标记鉴别猪瘟病毒疫苗毒株和流行毒株

建立一种检测CSFV的套式RT-PCR方法,旨在鉴别CSFV疫苗毒和流行毒.根据GenBank上公开发表的CSFV全基因序列,设计并合成了2对引物,建立了检测CSFV的套式RT-PCR方法,并对疫苗毒、SXYL株和SX01株扩增片段进行了序列测定和分析.结果表明,该方法检测CSFV cDNA含量的最低极限为3.4×10-5 ng/mL;特异性检测发现从PRRSV、SIV、PCV2和PRV阳性毒未扩增出特异性条带.序列分析表明,只有疫苗毒3′-NCR存在分子标记CTTTTTTCTTTT,SXYL株和SX01株均没有这一标记.建立的检测CSFV的套式RT-PCR方法灵敏度高,特异性强,是一种可行的检测方法.通过目的片段的序列分析,寻找分子标记CTTTTTTCTTTT,可以准确鉴别CSFV疫苗毒和流行毒.

Identify the Pandemic or Vaccine CSFV Strains by Molecular Markers

Establishment of a nested RT-PCR method for detection CSFV,the purpose of the sequenced fragments identified pandemic or vaccine CSFV.According to published on the GenBank CSFV gene sequence,designed and synthesized two pairs of primers,established a test method of CSFV by nested RT-PCR,amplification of vaccine,SXYL,and SXO1 strains fragment,analysis these sequences.The result of sensitivity showed that the most dilution cDNA of CSFV is 3.4×10^-5ng/mL; the result specialization showed that the method could not acquire any bandfrom PRRSV,SIV,PCV2 and PRV.Sequence analysis showed that only vaccine strain 3''-NCR has molecular markers CTTTTTTCTTTT,SXYL,and SXO1 strains had not it.The nested RT-PCR method of detection CSFV was high sensitivity and specificity,it was a viable method of detection.Through fragments of sequence analysis,looking for molecular markers CTTTTTTCTTTT,could accurately identify the pandemic or vaccine CSFV strains.