鸡传染性支气管炎病毒(M41)HI抗原的制备与初步应用

实验比较了不同方法制备IBV HI抗原,并比较不同PLC1处理浓度、不同保存时间等因素对制备IBV HI抗原的影响,筛选出制备抗原的最佳条件:选择10 U/ml的PLC1与等体积超速离心后的M41 IBV以及pH值7.40、.01 mol/L PBS稀释液混合,在37℃水浴作用2 h并不断振荡,取出后加入β-丙内酯4℃灭活24 h。在HI实验中,该抗原不与AIVH5、AIVH9、ND单因子血清反应,仅与IB阳性血清反应,有高度特异性。用高岭土37℃处理1 h可明显降低SPF阴性血清非特异反应。上述抗原在4℃保存3周血凝价达到最高。应用该抗原检测免疫前后鸡群IBV抗体,取得良好效果,证明用该方法与条件所制备的抗原可以用于IB诊断及临床实验。

Preparation and Preliminary Application in HI Antigen of Chicken Infectious Bronchitis Virus (M41)

Different preparation methods of IBV HI antigen were researched and the influence of different treated concentration of PLC1 and different storage time on preparation of IBV HI antigen were evaluated in this experiment.The optimal condition of antigen preparation was acquired.10 unit/ml of PLC1 was mixed with the same volume of IBV M41 diluted with pH 7.4 and 0.01 mol/L PBS after ultracentrifugation.The mixture was reacted in the water bath for 2 hours at 37 ℃ while being continually oscillated.Then Beta-C-lactone was added in it for 24 hours at 4 ℃ to inactivate IBV.The prepared antigen had high specificity and only reacted to IB positive serum,but not to AIVH5,AIVH9,ND and SPF chicken serum.The antigen was treated 1 hour at 37 ℃ by using china clay and its non-specific reaction to SPF chicken negative serum lowered significantly.Its HI titer reached to the maximum after keeping 3 weeks at 4 ℃.Application of this antigen in detecting the antibody to IBV of chicken indicated that it could be used in IB diagnosis and clinical trials.