新城疫病毒La Sota株和鸡传染性支气管炎病毒M41株低免疫鸡胚同胚培养研究

Inoculation and Culture in the Same Low-immunity Chicken Embryo of Newcastle Disease Virus La Sota Strain and Avian Infectious Bronchitis Virus M41 Strain

为研究新城疫病毒(NDV)La Sota株和鸡传染性支气管炎病毒(IBV)M41株在低免疫鸡胚中进行同胚接种培养的可行性,通过将不同稀释倍数的La Sota株和M41株接种同一低免疫鸡胚,分别测定收获尿囊液La Sota株和M41株含量,确定最佳的稀释倍数以及适宜的培养温度和培养时间。结果显示:La Sota株和M41株以10 000∶5 000的比例稀释,培养96 h病毒含量和收获量最佳,La Sota株含量可达到109.17EID50/0.1 m L,HA效价为1∶1 024,M41株含量可达到106.50EID50/0.1 m L。接种后孵育温度为36.5℃时,每胚收获量比37℃培养增加了1.0~1.2 m L。按照此工艺生产病毒制备疫苗免疫SPF鸡,NDV和IBV抗体均符合国家标准要求。试验探索并优化了La Sota株和M41株在低免疫鸡胚中同胚接种的培养条件,为大规模抗原生产提供数据支持。

In order to study the feasibility of inoculating the same low-immunity chicken embryos, Newcastle disease virus(NDV) La Sota strain and avian infectious bronchitis virus(IBV) M41 strain were used. La Sota strain and M41 strain with different dilutions were inoculated into the same low-immunity chicken embryo, La Sota and M41 content of the obtained virus solution was measured to determine the optimal dilution ratio and the appropriate culture temperature.The results showed that the virus content and harvested amount were the best when La Sota strain: M41 strain was diluted at a ratio of 10 000∶5 000 and cultured for 96 h. The La Sota content could reach 109.17 EID50/0.1 m L, HA titer was1∶1 024, and M41 content could reach 106.5 EID50/0.1 m L. The virus harvest per embryo at 36.5 ℃ was increased by 1.0 to 1.2 m L compared with that at 37 ℃. The vaccines were prepared by viruses produced according to this process and antibodies against NDV and IBV were complied with the national standards. The experiment explored and optimized the culture conditions of La Sota strain and M41 inoculated in the same low-immunity chicken embryo, and provided the data support for large-scale antigen production.