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| 一株2型牛病毒性腹泻病毒的分离鉴定及其在MDBK细胞上的克隆纯化 | |
| 引用本文： | 聂兆晶,田夫林,姜世金.一株2型牛病毒性腹泻病毒的分离鉴定及其在MDBK细胞上的克隆纯化[J].西北农业学报,2012,21(1):21-25. |
| 作者姓名： | 聂兆晶 田夫林 姜世金 |
| 作者单位： | 1. 山东农业大学动物医学院,山东泰安,271018 2. 山东省动物疫病预防与控制中心,济南,250022 3. 山东农业大学动物医学院,山东泰安271018;山东省动物生物技术与疫病防治重点实验室,山东泰安271018 |
| 基金项目： | 山东省自主创新成果转化重大专项计划 |
| 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。 |
| 关 键 词： | BVDV 分离鉴定 序列分析 TCID50 |
Isolation, Identification and Purification of a Genotype 2 Bovine Viral Diarrhea Virus in MDBK Cells |
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| NIE Zhaojing,TIAN Fulin and JIANG Shijin.Isolation, Identification and Purification of a Genotype 2 Bovine Viral Diarrhea Virus in MDBK Cells[J].Acta Agriculturae Boreali-occidentalis Sinica,2012,21(1):21-25. | |
| Authors: | NIE Zhaojing TIAN Fulin and JIANG Shijin |
| Institution: | 1,3(1.College of Veterinary Medicine,Shandong Agricultural University,Taian Shandong 271018,China; 2.Shandong Provincial Center for Animal Disease Control and Prevention,Jinan 250022,China;3.Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention,Taian Shandong 271018,China) |
| Abstract: | 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. |
| Keywords: | BVDV Isolation and identification Gene sequence analysis TCID50 |
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