摘要
以大肠杆菌表达的传染性法氏囊病病毒(IBDV)衣壳蛋白VP2致敏绵羊红细胞,建立间接血凝试验(IHA)。对参考阳性血清(鸡新城疫病毒抗血清、鸡传染性支气管炎病毒抗血清、鸡禽痘病毒抗血清)及鸡参考阴性血清等分别进行微量IHA,加上抑制试验,结果证实其特异性高。对商品肉用仔鸡的30个血清样品、IBDV重组亚单位疫苗免疫试验鸡的20个血清样品,分别进行微量IHA和琼脂免疫扩散试验(AGID),将肉用仔鸡血清的阳性检出率进行t检验(P<0.05);而20份免疫试验鸡血清样品IHA阳性检出率与AGID阳性检出率均为100%。对30份肉用仔鸡血清样品和20份免疫试验鸡血清样品检测得到的阳性血清的平均滴度进行方差分析,IHA的平均滴度与AGID的平均滴度之间均具有显著性差异。对20份免疫试验鸡血清及IBDV参考阳性血清以3个批次和第2批次IHA试验制剂,分别进行重复性试验,对其IHA平均滴度进行了方差分析,均无显著性差异。对20份免疫试验鸡血清以在4℃、保存期分别为3、6、9个月的IHA的试验制剂进行了微量IHA,检出的IBDV抗体平均滴度中,3个月的与6个月的无显著差异;3个月的与9个月的及6个月的与9个月的,均有显著差异,结果表明该IHA试验制剂在4℃的保存期可达6个月。
An indirect hemagglutination assay （IHA） was set up using sheep erythrocytes sensitized with capsid protein VP2 of infectious bursal disease virus （IBDV） which were expressed in Escherichia coll. Reference positive sera,such as chicken anti-sera to newcastle disease virus, chicken anti-sera to infectious bronchitis,chickens anti-sera to fowl pox virus, and reference chicken negative sera were detected with micro-IHA respectively,and inhibition test was carried out,and high specificity of the IHA was verified with experimental results. 30 sera samples of broilers and 20 sera samples of chickens vaccinated with IBDV recombinant subunit vaccine were detected by IHA and agar gel immunodiffusion test （AGID） respectively, and significant test to positive rate of broilers between IHA and AGID was analyzed with t test （P〈0.05）, but positive rate of broilers and chickens vaccinated were 100%. Significant test to mean antibody titer of positive sera of broilers and chickens vaccinated were analyzed with analysis of variance between IHA and AGID, and the test results were all P〈0.05. 20 sera samples of chickens vaccinated and IBDV reference positive sera were detected with 3 batches of and batch second of IHA experimental preparation in repeated test, mean antibody titer of broilers were analyzed with analysis of variance and the differences were no statistical significant. 20 sera samples of chickens vaccinated preserved 3,6,9 months at 4℃ were detected respectively with micro-IHA,and the difference of mean antibody titer was no statistical significant between 3 months and 6 months, but the differences of mean antibody titer were statistical significant between 3 months and 9 months,and between 6 months and 9 months. The experimental results made known that IHA experimental preparation was preserved to 6 months at 4℃.
出处
《湖北农业科学》
北大核心
2007年第2期271-275,共5页
Hubei Agricultural Sciences
基金
湖北省教育厅重点科技发展项目(992P0706)
