牛呼吸道合胞体病毒RT-PCR检测方法的建立及初步应用

Establishment of RT-PCR assay for detection of bovine respiratory syncytial virus

根据已发表的牛呼吸道合胞体病毒的全序列,选择保守性较强的N基因序列,利用Primer5.0设计合成特异性引物,建立能快速检测牛呼吸道合胞体病毒的RT-PCR方法。利用本方法对采集的38份临床症状类似的牛呼吸道合胞体病毒进行检测,检测结果中有10份为阳性。试验证明该方法具有很高的特异性及敏感性,能检测到1pg左右的病毒RNA,是一种快速有效的检测方法。

According to the published complete sequences of the Bovine Respiratory Syncytial Virus N gene,a pair of specific primers was designed and synthetized by using Primer5.0,then the RT-PCR method was established for detecting the Bovine Respiratory Syncytial Virus.Using the specific primers 287 bp band could be obtained from the BRSV by the method.Using the RT-PCR method clinical 38 samples,10 samples were positive.The result indicates that the RT-PCR is very high sensibility and specificity,it can detect 1 pg RNA of virus.So it is a quick and effective method for detecting the BRSV.