利用羊抗鸡抗体检测鹅血浆中的抗体水平  

Detection of Antibody Levels in Goose Serum Samples Using Goat-Anti-Chicken Secondary Antibody

作　　者：黄运茂[1] 苏记良[2] 于迎春[2] 李孝伟[2] 施振旦[2] 

机构地区：[1]仲恺农业技术学院生命科学学院,广东广州510225 [2]华南农业大学动物科学学院,广东广州510642

出　　处：《华南农业大学学报》2007年第2期95-98,共4页Journal of South China Agricultural University

基　　金：广东省自然科学基金项目(04205804);仲恺引进优秀人才科研启动项目(G2360232)

摘　　要：利用商品供应的山羊抗鸡抗体ELISA检测鹅血液中抗体水平的可行性.在相同条件下,以羊抗鸡抗体作为第2抗体,在ELISA分析中检测利用同一抗原免疫后鸡和鹅血浆样品中抗体水平.经过相同倍数梯度稀释的鸡和鹅血浆样品中,所测到的鹅血样的D450 nm值曲线在处理组和对照组均与鸡的D450 nm值曲线变化趋势相同,只是随血样的稀释倍数上升,鹅血样D450 nm值的下降速度较慢.结果表明只要反应条件适当,抗鸡抗体可以作为第2抗体用于ELISA检测鹅血液中的抗体.通过抗原/血浆浓度梯度交叉试验,ELISA中抗原的最适包被质量浓度为100μg/mL(150μL/孔),血样最佳稀释比为1∶12 800.ELISA测定所得鹅免疫试验中的抗体变化与理论预计结果一致,与在鸡上的变化也类似.The present study evaluated the feasibility and validity of using a biotinylated goat-anti-chicken secondary antibody in the ELISA for the detection of antibody titres in goose serum samples. Goose samples serially diluted produced a similar decay pattern of D450 nm values at a slightly slower rate than chicken samples. However, at dilution rates from 6 400 to 25 600, both chicken and goose samples yielded similar D450 nm values, which also exhibited maximal deviation from basal values of negative control sam- ples. Using goat-anti-chicken antibody as the secondary antibody in the ELISA, the optimal concentration of recombinant chicken inhibin fusion protein to coat the plate was determined to be 100 μg/mL （ 150 μL/well）. Using these criteria of plate coating and sample dilution rate, samples from geese immunized against recombinant chicken inhibin fusion protein produced D450 nm values, reflecting fluctuations of blood antibody levels in line with the theoretically changes after immunizations. The above results demonstrated that biotinylated goat-anti-chicken antibody could be used in ELISA to detect antibody levels in goose samples, provided that the samples be diluted at the rate between 6 400 and 25 600.

关 键 词：抗鸡抗体 鹅 抗体检测 酶联免疫吸附试验(ELISA) 

分 类 号：S852.4[农业科学—基础兽医学]