摘要
参照多株不同基因型和基因亚型牛病毒性腹泻病毒(BVDV)5′UTR相对保守的基因序列,设计3对引物,其中1对能够扩增不同基因型BVDV 5′UTR片段(288 bp),另2对分别能够扩增BVDV-1型和BVDV-2型5′UTR片段(195 bp和116 bp)。扩增片段大小与预计片段一致。对PCR反应条件进行优化,建立了一种特异、敏感的RT-nPCR检测方法。敏感性试验表明,敏感性为10-2TCID50/mL。特异性试验表明,对猪瘟兔化弱毒疫苗株、牛轮状病毒、牛传染性鼻气管炎病毒的检测结果均为阴性。通过对新疆部分地区无BVD临床症状的牛场208份奶牛血样检测,阳性率为50.43%,表明该方法具有快速、简便、特异性强和灵敏度高等特点,可以作为BVDV隐性感染临床诊断的有效方法。
According to 5′UTR sequence of different genotypes and subgroups of Bovine viral diarrhea virus（BVDV） strains,three pairs of primer were designed.One of them can amplify 5′UTR（288 bp） of all BVDV genotypes.Another two primers were specific for BVDV-1 and BVDV-2（195 bp and 116 bp）,respectively.The length of amplified fragment was consistent with prediction.PCR conditions were optimized.A RT-nPCR method was established for detecting BVDV.10-2 TCID50 of BVDV per 100 mL could be detected.Hog cholera lapinized virus（HCLV）,bovine rotavirus（BRV）,infectious Bovine Rhinotracheitis Virus（IBRV） were negative in specific experiment.208 clinical samples were detected using the RT-nPCR method with positive percentage of 50.43.The results indicated that the RT-nPCR is very specific and sensitive.So it would be an available method for detecting the silent infection of BVDV.
出处
《西北农业学报》
CAS
CSCD
北大核心
2010年第10期6-9,15,共5页
Acta Agriculturae Boreali-occidentalis Sinica
基金
"973"计划前期研究专项项目(2008CB117017)
兵团博士资金项目(2007JC15)
