应用斑点免疫金染色法检测猪细小病毒病抗体的研究

Detection of the serum antibody against porcine parvovirus by the dot immuno-gold staining technique

本研究初步建立了斑点免疫金染色法 (Dot- IGS)检测 PPV抗体的方法。试验采用经蔗糖密度梯度离心制备的 PPV提纯抗原和 p H7.4、直径 5 0～ 6 3.5 mm的胶体金来检测 PPV抗体 ,取得了较为满意的效果 ;该法对猪细小病毒病抗体最小检测量为1.0 0 8× 10 - 1 0 g/ ml,灵敏性高 ;不同猪布鲁氏菌病、猪伪狂犬病、猪衣原体病、猪口蹄疫、猪瘟、猪弓形体病等的阳性血清出现交叉反应 ,特异性强且重复性好。另该法还具有胶体金制备容易、操作简单快捷等优点 ,是对 PPV抗体检测的重要方法。

A Dot Immun-gold staining (Dot-IGS) new way was developed for the detection of porcine parvovirus (PPV) antibody. It was satisfactory Results that purified PPV antigen obtained by sucrose density gradient centrifuging and the colloidal gold with pH of 7.4?diameter of 50～63.5 mm were used to detect the PPV antibody. The detection limit of PPV antibody by the Dot-IGS was 1.008×10~ -10 g. It sshows that the sensitivity of Dot-IGS was high. A cross-reaction tests with positive sera of brucellosis, swine pseudorabies, swine chlamydiosis, swine foot and mouth disease, swine toxoplasmosis and swine fever demonstrated the high specificity of the Dot-IGS, no cross-reaction occurred. Much advantages of Dot-IGS for detection of PPV antibody was described: colloidal gold made easily, the experiment is easy and rapid and the results are easy to read. The development of Dot-IGS to detect PPV antibody is an important new way.