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目的:为了分析SEREX抗原克隆在异体食管癌患者血清中和健康对照人血清中的反应情况,我们发展了一种基于免疫印迹的高通量肿瘤相关抗原筛选和鉴定方法。方法:采用新发展的基于免疫印迹原理的抗原克隆微阵列检测方法与传统的噬菌斑检测法对食管癌患者血清中IgG的反应情况进行了比较,并采用两种方法分析了两个SEREX阳性克隆与40例食管癌患者血清和40例健康人血清中的IgG的阳性反应率。通过ELISA对两种方法得到的结果进行了验证。结果:两个阳性克隆在两种检测方法中均与同一例食管癌病人血清起反应,采用抗原克隆微阵列检测方法检测p53和rps3两个克隆阳性率分别为25%和37.5%,采用噬菌斑检测法检测p53和rps3两个克隆阳性率分别为27.5%和37.5%;ELISA方法检测p53和rps3两个克隆阳性率分别为25.0%和31.25%。结论:本研究采用两种方法检测患者血清IgG反应的结果是一致的,抗原克隆微阵列检测方法可以代替传统的噬菌斑筛选法。
OBJECTIVE: To develop a serum-based screening and identification of high-throughput tumor-associated antigens based on immunoblotting in order to analyze the response of SEREX antigen clones in serum of patients with esophageal cancer and healthy controls. Methods: The newly developed immunoblotting-based antigen clone microarray assay was compared with the traditional plaque assay for serum IgG in patients with esophageal cancer and two serological positive samples were analyzed by two methods The positive rate of clonality of IgG in the sera of 40 esophageal cancer patients and 40 healthy volunteers. The results of the two methods were validated by ELISA. Results: The two positive clones reacted with the same serum of esophageal cancer patients in both detection methods. The positive rates of p53 and rps3 were 25% and 37.5%, respectively, using phage clone microarray detection method. The positive rates of p53 and rps3 were 27.5% and 37.5%, respectively. The positive rates of p53 and rps3 by ELISA were 25.0% and 31.25% respectively. CONCLUSION: The results of this study using two methods to detect serum IgG in patients are consistent. Antigen clone microarray assay can replace traditional plaque screening method.