用分子标记鉴别猪瘟病毒疫苗毒株和流行毒株

Identify the Pandemic or Vaccine CSFV Strains by Molecular Markers

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3.4×10-5ng/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。

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 SX01 strains fragment, analysis these sequences. The result of sensitivity showed that the most dilution cDNA of CSFV is 3.4×10^-5 ng/mL; the result specialization showed that the method could not acquire any band from PRRSV, SIV, PCV2 and PRV. Sequence analysis showed that only vaccine strain 3＇-NCR has molecular markers CTTTTTTCTTTT, SXYL and SX01 strains had not it. The nested RT-PCR method of detection CS- FV 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.