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发明公开
CN108152498A 一种适用于牛BVDV、IBRV、PIV3、BRSV灭活疫苗的灭活检验方法 无效 - 驳回

一种适用于牛BVDV、IBRV、PIV3、BRSV灭活疫苗的灭活检验方法
摘要：
本发明公开了一种适用于牛BVDV、IBRV、PIV3、BRSV灭活疫苗的灭活检验方法。本发明涉及生物医药领域，具体涉及一种针对牛病毒性腹泻病毒、牛传染性鼻气管炎病毒、牛副流感病毒、牛呼吸道合胞体病毒灭活疫苗的灭活检验方法，包括以下步骤：1）灭活后的病毒液在适宜细胞上吸附40-60min，期间振摇2-3次；2）吸附后，在细胞转瓶中加入一定体积的细胞维持液继续培养，观察细胞病变，对于无细胞病变的病毒液，收获细胞培养液，并冻融2-3次；3）取冻融后的细胞培养液10-15ml，接种于适宜细胞上吸附，继续培养。进行观察和荧光检测，如没有特异性荧光，则表明灭活完全。本发明中公开的技术方案保证了抗原浓度和病毒感染时间。经本发明灭活检验方法检验病毒的检出率高，可以有效确保灭活疫苗的生物安全。
摘要（英）：

The invention discloses an inactivation inspection method suitable for BVDV, IBRV, PIV3 and BRSV inactivated vaccines, and relates to the field of biological medicines, in particular to an inactivation inspection method for BVDV, IBRV, PIV3 and BRSV inactivated vaccines. The inactivation inspection method comprises the following steps: (1) adsorbing an inactivated virus liquor on an appropriate cell for 40-60 min, and shaking for 2-3 times in the period; (2) after adsorption, adding a certain volume of a cell maintenance fluid in a cell spinner bottle, continuously performing culture, observing cytopathy, as for the virus fluid free from cytopathy, harvesting a cell culture fluid, and performing freeze thawing for 2-3 times; and (3) taking 10-15 ml of the frozen and thawed cell culture fluid, inoculating the taken cell culture fluid on the appropriate cell for adsorption, and continuously performing culture. Observation and fluorescence detection are performed, and if no specific fluorescence appears, complete inactivation is manifested. According to the technical scheme disclosed by the prevention, the anti-gen concentration and the virus infection time are guaranteed. The virus detection rate of the inactivation inspection method is high, and the bio-safety of the inactivated vaccines can be effectively ensured.

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