目前检测卵黄抗体的方法很多,但IHA方法具有简便快速的优点,本研究的目的是为了建立抗犬瘟热高免卵黄抗体检测方法。采用犬瘟热病毒和犬细小病毒二联疫苗处理作为疫苗抗原,用犬瘟热病死犬的肝脏部分处理后作为组织抗原,制备敏化绵羊红细胞,用IHA方法确定了犬二联疫苗抗原和犬瘟热病毒组织抗原的最适含量。二联疫苗提取抗原的含量为0.7 mg/mL致敏红细胞时,与犬瘟热单克隆抗体反应显著,抗体效价为1∶256;而患犬瘟热松狮犬的肝脏组织抗原蛋白浓度为0.5mg/mL,与犬瘟热单克隆抗体反应显著,抗体效价为1∶128。经过IHA验证,制备的松狮犬瘟热肝组织抗原,蛋白含量0.5 mg/mL 1%致敏红细胞,对应的单抗、鸡血清均出现了凝集,阳性对照也出现凝集,阴性对照未出现凝集。建立的IHA抗体检测方法,为准确检测制备抗犬瘟热高免卵黄抗体的效价奠定了基础。%The purpose of this study was to establish a method to detect hyperimmunized immunoglobuin of Yolk of canine distemper virus. Canine distemper virus and canine parvovirus treatment bivalent vaccine was used as the vaccine antigen, and the liver of a dead canine caused by canine distemper virus was treated and used for tissue antigen. The sensitized sheep red blood cells were prepared, The IHA method was applied to determine the optimal levels of canine bivalent vaccine antigen abd distemper virus tissue antigens. When extraction of bivalent vaccine antigen content was 0.7 mg / mL sensitized red blood cells, the monoclonal antibody reacted with canine distemper significantly, and the antibody titer was 1：256;The liver tissue antigen concentration of Chow Chow dogs suffering from distemper was 0.5 mg / mL, the monoclonal antibody reacted with canine distemper significantly, and the antibody titer was l：128.After IHA verification, protein content was 0.5 mg/ roLl% sensitized red blood cells in liver tissue of antigen Chow suffering from distemper,the corresponding monoclonal antibody and chicken serum agglutination occured, positive control agglutination also occured, agglutination negative control did not appear . The establishment of an IHA method can provide a basis for detecting hyperimmunized immunoglobuin of Yolk of canine distemper virus.
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