摘要
制备猪瘟荧光抗体并用于猪只活体扁桃体组织抹/涂片检测。将制备的猪瘟高免血清采用硫酸铵分级沉淀法纯化免疫球蛋白,反向透析法标记荧光素,过葡聚糖凝胶柱(Sephadex G-50)去除游离荧光素,通过扁桃体组织匀浆吸附去除异嗜性或交叉反应抗体。以5%FCS做稀释液,以RT-PCR检测阳性样本的冰冻切片确定其最大稀释度为1/60,最佳工作浓度为1/30,阻抑试验证明荧光为特异荧光。用标记的荧光抗体检测活体扁桃体样本39份,检出阳性11份,与IDEXX猪瘟抗原检测试剂盒复核结果完全吻合。结果表明,本实验所制备的猪瘟荧光抗体具有很高的灵敏性和特异性,可用于猪瘟净化中扁桃体组织抹/涂片检测。
To prepare fluorescent antibody of classical swine fever （CSFV） used in the tonsil tissue detection of living swines, immunoglobulin was extracted from hyper - immunized serum by ammonium sulphate step salting out, while fluorescein isothioeyanate （FITC） was marked by retrodialysis, the redundant liberation FITC was removed by sephadex G50, and the cross - reacting or the isotropic antibody was further removed by the adsorbing of tonsil homogenate. The CSFV positive tonsil proved by RT-PCR was made into frozen section to determine the final level diluted by 5% FCS. The working concentration was 1/30 and the final dilution 1/60, depression test proved the fluorescence were special. The fluorescent antibody was utilized in the clinical test of classical swine fever in 39 tonsils from different places and 11 tonsils were screened as positive, the result was proved by IDEXX CSFV Ag Test Kit. The results showed that the precipitated CSFV fluorescent antibody had high sensitivity and specificity and could be used in the clinical CSFV depuration of tonsil tissue detection.
出处
《中国兽药杂志》
2008年第12期16-19,共4页
Chinese Journal of Veterinary Drug
关键词
猪瘟
荧光抗体
制备
应用评价
classical swine fever
fluorescent antibody
preparation
applied evaluation
