外源病毒检验用抗鸡传染性支气管炎病毒特异性血清的制备与检定

Preparation and Identification of Specific Antiserum against Infectious Bronchitis Virus Used for Exogenous Virus Test

为制备外源病毒检验用抗鸡传染性支气管炎病毒(IBV)特异性血清,对300只3~4周龄的SPF鸡进行基础免疫和加强免疫;最后一次免疫21 d后,采血并分离血清;对血清经混合、分装、冷冻、真空干燥后,进行无菌检验、外源病毒检验、剩余水分测定、中和效价测定和特异性检验。结果表明:本研究制备的抗IBV特异性血清的无菌检验、外源病毒检验和剩余水分测定结果均符合现行《中华人民共和国兽药典》规定;血清中和效价为1:800,与IBV H120种毒株的中和指数为106.3,与鸡传染性法氏囊病毒B87株、鸡新城疫病毒La Sota株、鸡传染性喉气管炎病毒、鸡痘病毒、禽呼肠孤病毒S1133株的中和指数均不大于10;通过AGP、ELISA、HI等试验,确定该血清中不含其他禽源外源病毒抗体。本研究为鸡传染性支气管炎活疫苗或毒种的外源病毒检验提供了具有良好特异性和高效价的中和用血清。

In order to prepare the specific antiserum against Infectious bronchitis virus（IBV）for exogenous virus test,basic and strengthened immunization were conducted towards 300 SPF chickens at the age of 3~4 weeks. Twenty-one days after the last immunization,the blood were collected and serum samples were isolated. After being mixed,packaged,frozen and dried up by vacuum,the serum samples were examined by sterility test,exogenous virus test,residual moisture test,neutralization titer determination and specificity test. The data results of sterility test,exogenous virus test and residual moisture test complied with the specifications of China Veterinary Pharmacopoeia. The neutralization titer of the serum was 1：800,and the neutralization index of the antiserum neutralizing with IBV H120 could reach to 106.3,while the values were below 10 when neutralizing with Infectious bursal disease virus（B87 strain）,Newcastle disease virus（La Sota）,Avian infectious laryngotracheitis virus,Fowlpox virus,and Avian reovirus. Besides,there was no antibody against other avian disease viruses in this serum according to APG,ELISA and HI test. In conclusion,antiserum samples with high specificity and neutralization titer were successfully prepared for detecting the exogenous virus mingled in IBV live vaccines and seed virus.