猪瘟病毒和牛病毒性腹泻病毒双重RT-PCR方法的建立和初步应用

根据GenBank上已发表的猪瘟病毒(CSFV)和牛病毒性腹泻病毒(BVDV)的全基因序列,进行对比分析,分别设计合成两对能特异性扩增CSFV、BVDV的引物。经过条件优化后,建立了检测CSFV和BVDV的双重RT-PCR方法,扩增两种病毒的片段,大小分别为938、650 bp。应用该方法对11批牛睾丸细胞、7批胎牛血清、60个批次的猪瘟细胞苗、10份全血样及10份组织样进行检测。通过试验证明,所建立的方法具有良好的特异性和敏感性,为防止猪瘟细胞苗的污染及进行CSFV和BVDV鉴别诊断提供了有效方法。

Establishment and Initially Application of Double RT-PCR Detection Method for Both Classical Swine Fever Virus and Bovine Viral Diarrhea Virus

According to the complete genome sequences of classical swine fever virus (CSFV) and bovine viral diarrhea virus (BVDV) in GenBank, two pairs of primers were designed and synthesized. A double RT-PCR was developed based on these two pairs of primers which amplified the CSFV virus-specific segment with 938 bp and the BVDV virus-specific segment with 650 bp in sizes after the conditions of PCR were optimized. Approved this method of 11 batches of bovine testicular cells,7 batches of fetal bovine serum,60 batches of swine fever vaccine,10 full-blood and 10 tissue samples for testing. The experiment showed that the developed method has good specificity and sensitivity of cell vaccine to prevent the pollution of BVDV and the diagnosis of CSFV and BVDV provides an effective method.