鸡传染性支气管炎病毒血凝抑制试验抗原的研制

为应用血凝抑制(HI)试验检验鸡传染性支气管炎疫苗免疫效力(血清、卵黄抗体水平),建立了鸡传染性支气管炎病毒HI试验抗原的制备方法。该方法是通过选取抗原谱最广的鸡传染性支气管炎病毒(IBV)M41株,经SPF鸡胚增殖培养36h后无菌收取鸡胚尿囊液,4℃、12 000r/min离心10min,上清用聚乙二醇(PEG)20000浓缩100倍;兔源A型产气荚膜梭菌中国标准株(C57-1株)37℃增殖培养18h,4℃、12 000r/min离心10min取上清,经PEG 20000透析袋浓缩5倍后通过0.20μm滤膜过滤除菌,然后将IBV液和A型产气荚膜梭菌菌液(含α毒素)二者按一定比例混合后经37℃恒温振荡感作2h,4℃经48h后制成。经大量试验表明,制备的IBV HI试验抗原效价高、稳定性好,可替代进口抗原应用于鸡群IBV疫苗免疫后血清抗体及卵黄抗体的HI效价检测。

Development of HI Test Antigen of Avian Infectious Bronchitis Virus

In order to determine the serum antibody level of avian infectious bronchitis virus (IBV)in the inactivated vaccine,the preparation method of HI test antigen of IBV was established.The widest spectrum of IBV M41 strain was selected and proliferated in allantoic fluid of SPF chick embryos for 36 h,and then centrifuged in 4℃ and 12 000 r/min for 10 minutes,and last concentrated 100 fold by the polyethylene gly-col (PEG)20000 dialysis bag.Rabbit Clostridium perfringens type A standard Chinese strain (C57-1 strain)was first cultured for 18 h,then centrifuged supernatant in 4℃ and 12 000 r/min for 10 minutes, and then concentrated 5 fold by the PEG20000 dialysis bag,and last filtered by 0.20 μm membrane.The IBV HI antigen was made by mixing the IBV virus fluid and type A Clostridium perfringens liquid (con-taining alpha toxin)according to a certain proportion,then by the constant temperature of 37℃ for 2 h,and then in 4℃ for 48 h.It was showed that the prepared IBV HI test antigen had high titer and good stability, and could successfully replace imported antigen by using for the HI detection of serum antibody and egg yolk antibody in the sera of chickens immunized with IBV vaccine by a large number of experimental re-sults.