嗜肺巴氏杆菌外膜蛋白和脂多糖抗原在血清学诊断中的意义

The Significance of Outer Membrane Protein and Lipopolysaccharide of Pasteuralla pneumotropica in Serological Diagnosis

目的 评价嗜肺巴氏杆菌外膜蛋白 (OMP)和脂多糖 (LPs)作为血清学诊断抗原的敏感性和特异性。方法 用OMP、LPS和全菌 (WC)作为Westernblot和ELISA的诊断抗原检测自然感染和实验感染嗜肺巴氏杆菌小鼠相应的IgG抗体滴度 ,同时测定 3种抗原与实验动物常见致病菌的交叉反应。结果 与嗜肺巴氏杆菌自然感染和实验感染小鼠血清的ELISA反应中 ,不同时期 ,LPS作为诊断抗原时血清抗体阳性率最高 ,WC次之 ,OMP最低。自然感染小鼠群中 ,出生 4周LPS抗体阳性率即可达 80 % ,而同期的WC和OMP仅为 2 5 %和 2 0 % ,故LPS敏感性最高。与实验动物常见致病菌免疫血清和阴性种鼠血清的ELISA反应中 ,WC抗原表现出较高的吸光度 (A)值 ,经Westernblot证实 ,其反应为非特异性反应 ,LPS抗原特异性最强 ,OMP抗原次之。结论 混合多株具有型或种特异性的OMP或LPS作为ELISA的诊断抗原 ,无论从特异性和敏感性上均高于全菌抗原。

Objective To compare the sensitivity and specificity of outer membrane protein (OMP) and lipopolysaccharide (LPS) of Pasteuralla pneumotropica used as serological diagnostic antigens to detect Pasteuralla pneumotropica.Methods LPS,OMP and whole cell(WC) were used as ELISA and Western Blot antigens to characterize the IgG antibody of mice with natural and experimental infection of Pasteuralla pneumotropica, meanwhile the cross_reaction of three antigens to common laboratory animal pathogens was also evaluated by ELISA and Western Blot methods.Results Three antigens were tested with sera from experimentally and naturally infected mice in different ages via ELISA.LPS showed the greatest antibody ratio,WC antigen showed lower,and OMP lowest.In naturally infected mice colony,the antibody ratio of LPS in 4_week age mice was 80%,while that of whole cell antigen and OMP only 25% and 20% respectively,showing that LPS was the most sensitive one.Three kinds of antigens were also tested with sera from uninfected mice and antisera of common laboratory pathogens via ELISA and Western Blot.WC antigen had a high A value in ELISA,but it was proved to be unspecific using Western Blot.LPS was the most specific antigen,and OMP second.Conclusion The mixed species specific and typical specific OMP and LPS in ELISA to detect latent infection of P.pneumotropica in laboratory animals have more specific and sensitive than WC antigen.