鸡传染性支气管炎病毒H120株和H52株种毒的制备与鉴定

Preparation and Identification of Chicken Infectious Bronchitis Virus H120 and H52 Strain Seeds

鸡传染性支气管炎病毒H120株、H52株种毒由中国兽医药品监察所制备和供应,种毒的保存期为10年,为了保证种毒的质量和供应,需要定期制备种毒。将H120株、H52株毒种接种SPF鸡胚进行传代培养,收获鸡胚尿囊液,与50 g/L牛乳蔗糖溶液等体积混匀,进行分装和冷冻真空干燥。对其进行剩余水分测定、真空度测定、病毒含量、安全性、免疫原性、纯净(无菌检验、支原体检验、外源病毒)和特异性检验,检验结果表明,各项检验均符合规定。同时为了区分H120株、H52株种毒,用实验室已建立的RT-PCR和酶切方法对两种种毒进行鉴别检验,结果表明能准确区分H120株和H52株种毒。试验结果为疫苗生产提供了能长期保存且稳定性好、高滴度的生产用种毒,也可为其他禽源病毒种毒的制备提供借鉴。

The chicken infectious bronchitis virus(IBV)H120,H52 strain seeds were prepared and supplied by China Institute of Veterinary Drug Control.The preservation period of the seeds were 10 years.In order to ensure the quality and supply,we need to prepare the seeds regularly.H120 and H52 strain seeds were inoculated in SPF chicken embryos for subculture.Allantoic fluid was harvested and well-mixed with the equal volume 50 mL/L milk sucrose solution.After separation and lyophilization,the allantoic fluid was used for residual moisture test,vacuum degree,virus content determination,safety,immunogenicity,sterility test,mycoplasma test,exogenous virus test and specificity test.All the results showed that the lyophilizated seeds meet the demand of regulation.While molecular biology identification could distinguish between IBV strains H120 and H52.The research provided seeds which could be kept for long term,showed high stability and high titer for vaccine production,also provided the reference for the preparation of other poultry virus seeds.