间接ELISA检测雏鸡烟曲霉菌抗体的研究

Detection of Antibody to Aspergillus Fumigatus in Sera of Chicks with Indirect Elisa

以烟曲霉菌丝提取物作为包被抗原 ,建立了检测雏鸡血清中烟曲霉菌抗体的间接 ELISA方法。特异性试验和重复性试验证明该方法特异性高 ,重复性好 ,其抗原包被浓度为 1 2 4μg/ ml,血清最适稀释度为1∶ 1 6,抗原最佳包被时间为 4℃过夜 ,血清反应时间为 3 7℃作用 1 .5 h。对 1 2 0份人工感染烟曲霉菌的雏鸡血清试验发现 ,间接 ELISA检出时间比琼脂扩散试验早 7d以上 ,检出率高约 40 .8%。表明间接

The indirect ELISA with coating mycelial extract antigen of A. fumigatus was established to detect specific antibody to A. fumigatus in sera of chicks and the assay was confirmed to be specific and reproducible. The concentration of coating antigen was 124 μg/ml and that of serum samples was 1∶16. The optimum coating time is a night at 4℃.Serum reaction time is 1.5 h at 37℃. Experimentally infected chicks were assayed for specific antibody by indirect ELISA and immunodiffusion (ID) methods at the same times and the sensitivities of two methods were compared. The result showed that the positive time of ELISA test was more than 7 days earlier than that of ID and the positive rate was 40.8% higher . All the results further confirmed that indirect ELISA test is a specific and sensitive method to detect A. fumigatus antibody.