摘要
根据GenBank中登录的牛病毒性腹泻病毒(BVDV)5′UTR基因序列,设计合成了1对特异性引物,建立了BVDV的RT-PCR检测方法。该方法从BVDV标准毒株NADL中扩增出了279bp的特异性片段,而对猪瘟病毒、猪繁殖与呼吸综合征病毒、牛传染性鼻气管炎病毒、牛轮状病毒、MDBK细胞等的扩增结果均为阴性。经对标准毒株的细胞毒进行检测,其敏感度达10-1×TCID50。提取9份细胞毒总RNA后,进行一步法RT-PCR扩增可获得一致的检测结果,说明该方法重复性很好。应用该方法对45份临床疑似患牛的样品进行检测,结果检出4份阳性,表明该方法具有较好的临床应用性,且检测结果与参考实验室检测结果完全相符,可用于BVDV的快速诊断。
A reverse transcription PCR(RT-PCR)for detection of bovine viral diarrhea virus(BVDV)was established using a pair of specific primers based on the 5'-untranslated region gene of bovine viral diarrhea virus(BVDV)published in GenBank.This method specifically amplify a 279bp of fragment from BVDV NADL strain,but not from classical swine fever virus,porcine reproductive and respiratory syndrome virus,infectious bovine rhinotracheitis virus,bovine rotavirus and normal MDBK cells.The results show that the sensitivity of the one-step RT-PCR method was 0.1×TCID50.In a test of 45 samples suspected with bovine disease,the result shows 4 samples were positive with BVDV.The established RT-PCR was specific,sensitive,efficient and rapid for the detection.
作者
张康
王丹阳
王旭荣
张凯
张景艳
王磊
李建喜
ZHANG Kang;WANG Dan-yang;WANG Xu-rong;ZHANG-Kai;ZHANG Jing-yan;WANG Lei;LI Jian-xi(Lanzhou Institute of Husbandry and Pharmaceutical Sciences,Chinese Academy of Agricultural Sciences,Gansu Province Veterinary Drug Engineering and Technology Research Center,Lanzhou 730050)
出处
《中国奶牛》
2018年第10期32-35,共4页
China Dairy Cattle
基金
国家奶牛产业技术体系科学家岗位项目(CARS37-06)
公益性行业(农业)科研专项(201303040-01)
作者简介
张康(1987-),男,甘肃兰州人,硕士,主要从事奶牛疾病研究工作;通讯作者:李建喜(1971-),男,研究员,博士生导师。
