摘要
为了解牛病毒性腹泻病毒(BVDV)在猪用活疫苗生产过程中的污染情况,利用抗原捕获ELISA方法对用于制作原代睾丸细胞的犊牛血清进行检测。将检测结果为阳性的1份犊牛血清,接种于MDBK细胞上,盲传3代,出现明显细胞病变。用BVDV 5′-UTR端特异性引物进行RT-PCR扩增,并对扩增片段进行克隆测序和序列分析。结果表明,分离得到一株2型牛病毒性腹泻病毒,并将其命名为JN-2。将该病毒用蚀斑方法进行克隆纯化,并对纯化前后病毒的TCID50进行测定比较,结果发现纯化后病毒滴度显著提高。
In order to study the contamination status of Bovine viral diarrhea virus（BVDV） on the process of live vaccine production for swine,the calf blood serum which served as primary cell was detected by antigen capture ELISA.The positive serum was inoculated on MDBK cells and reproduced blindly for 3 generations,and then the virus caused cells pathological changes.Through the RT-PCR and the gene sequencing,a BVDV of genotype 2 named JN-2 was obtained.By plaque assay and TCID50 detection,the JN-2 isolate was purified and its viral titer rose significantly in MDBK cells.
出处
《西北农业学报》
CAS
CSCD
北大核心
2012年第1期21-25,共5页
Acta Agriculturae Boreali-occidentalis Sinica
基金
山东省自主创新成果转化重大专项计划(2008ZHZX1A1101)
