牛呼吸道合胞体病毒RT-PCR检测方法的建立

Establishment of RT-PCR for Detection of Bovine respiratory syncytial virus

根据GenBankTM上发表的牛呼吸道合胞体病毒核衣壳蛋白N基因序列,设计合成了一对特异性引物,扩增大小为596bp的目的片段,通过特异性试验、敏感性试验和重复性试验建立了牛呼吸道合胞体病毒的RT-PCR检测方法。所建立的牛呼吸道合胞体病毒的RT-PCR方法与牛腺病毒、牛副流感病毒、牛传染性鼻气管炎病毒、牛无浆体均无交叉反应,该方法的敏感性可达1TCID50。结果表明,该方法具有快速、敏感、特异性强和重复性好等特点,可作为牛呼吸道合胞体病毒检测的一种方法。

Specific primers were designed and synthesized according to the reported nucleocapsid （N） protein gene seguence from bovine respiratory syneytial virus （BRSV） in GenBankTM. The size of 596 bp fragment was amplified by different condition optimization, then the RT-PCR diagnostic method of BRSV was con- structed by the specific,sensitive and reproducible assays. The diagnostic method of BRSV was no crossreaction with the closely related clinical symptom pathogen, such as bovine adenovirus, bovine parainflu- enza virus, bovine infectious rhinotracheitis virus, Anaplasrna rnargina ; the sensitivity of this method was at least 1 TCID50. The results of a series of tests indicated that it was a rapid,sensitive, specific, reproducible method, and could be used as a method of detection for bovine respiratory syncytial virus infection.