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目的:建立检测抗P53蛋白抗体的ABC-ELISA法,并进行初步临床应用的总结。方法:用人工合成P53蛋白N端25 个氨基酸肽链为抗原,建立了敏感的亲和素-生物素ELISA法,240份肿瘤患者和100份正常人血清抗P53 蛋白抗体,用免疫组化检测肿瘤细胞内P53蛋白表达情况。结果:肿瘤患者血清抗P53 蛋白抗体阳性率分别为:结直肠癌22% 、肺癌34% 、乳腺癌25% 、胃癌36% 、原发性肝癌32% ;而正常人血清中抗P53 蛋白抗体的阳性率≤2% 。对20份肿瘤组织进行免疫组化检测,肿瘤细胞内P53 蛋白阳性率为60% 。此外,用Pharm a-Cell试剂盒检测90 份结直肠癌患者和乳腺癌患者血清,结果与ABC-ELISA 法无统计学差异(P> 0.05)。结论:抗P53 蛋白抗体在肿瘤患者中检出率显著高于正常人群,它可以作为一种新的肿瘤标志物,对肿瘤的诊断起到辅助作用。
OBJECTIVE: To establish an ABC-ELISA method for detecting anti-P53 protein antibodies, and to summarize the preliminary clinical application. METHODS: Using a synthetic 25 amino acid peptide chain at the N-terminus of P53 protein as an antigen, a sensitive avidin-biotin ELISA was established. 240 patients with tumor and 100 normal human serum anti-P53 protein antibodies were detected by immunohistochemistry. P53 protein expression in tumor cells. Results: The positive rates of serum anti-P53 protein antibody in tumor patients were: colorectal cancer 22%, lung cancer 34%, breast cancer 25%, gastric cancer 36%, primary liver cancer 32%; and normal human serum anti-P53 protein antibody Positive rate ≤ 2%. The immunohistochemical detection of 20 tumor tissues showed that the positive rate of P53 protein in tumor cells was 60%. In addition, using the Pharm a-Cell kit to detect the serum of 90 patients with colorectal cancer and breast cancer, the results were not statistically different from the ABC-ELISA (P> 0.05). Conclusion: The detection rate of anti-P53 protein antibody in tumor patients is significantly higher than that in the normal population. It can be used as a new tumor marker and can help to diagnose tumors.