猪流行性腹泻病毒敏感细胞的筛选方法及应用

Screening Method and Application of Porcine Epidemic Diarrhea Virus Sensitive Cells

关键词:  猪流行性腹泻  克隆细胞  病毒含量

Key words:  Porcine epidemic diarrhea  Clone cells  Virus content

摘要:

为了提高猪流行性腹泻抗原的病毒含量，对敏感Vero细胞的筛选方法及应用进行了研究。从6株不同来源的Vero细胞中筛选出敏感细胞进行单细胞克隆，对每个克隆细胞进行猪流行性腹泻病毒培养实验，筛选出病毒含量最高的克隆细胞，即为敏感克隆细胞。对敏感克隆细胞进行猪流行性腹泻病毒感染实验，当细胞病变达到80%以上时冻融收获，病毒含量能达到107.5 TCID50/mL。试验表明，本研究克隆出了一株对猪流行性腹泻病毒最敏感的VERO-D细胞，生产出的抗原病毒含量比常规工艺提高0.5个滴度以上。制备的成品检验结果和常规产品无差异，且免疫效果良好，为猪流行性腹泻灭活疫苗生产工艺的优化提供了数据支持。

Abstract:

In order to increase the virus content of porcine epidemic diarrhea antigen, the screening method and application of sensitive Vero cells were studied. Sensitive cells were selected from a pool of six Vero cell lines originating from diverse sources for single-cell cloning. Porcine epidemic diarrhea virus culture experiments were conducted on each cloned cell, and subsequent screening identified the clone cells exhibiting the highest viral content, hereby referred to as sensitive clone cells. Then, the sensitive cloned cells were infected with porcine epidemic diarrhea virus, freeze-thaw harvest was conducted when the cytopathic effect reaches more than 80%, and the virus content could reach 107.5 TCID50/mL. The results show that the VERO-D cell, which is most sensitive to porcine epidemic diarrhea virus, was successfully cloned in this study, and it can produce antigen content 0.5 titer higher than the conventional process. Besides, the inspection results of the prepared product showed no difference compared to those of the conventional product, and the immunization effectiveness was excellent. These findings provide data support for the optimization of the production process of porcine epidemic diarrhea inactivated vaccine.