检测抗P53蛋白抗体ABC-ELISA法的建立和初步临床应用

Development of ABC-ELISA method for detecting anti P53 antibodies and its primary clinical application

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 develop ABC ELISA for detecting antibodies of P53 protein (P53 Abs) and evaluate the significance of P53 with the pathalogical stage and the prognosis of tumor. Methods: Serum P53 Abs were detected in 240 tumor patients and 100 normal adults by ABC ELISA. Expression of P53 in the solid tumor was examined by immunohistochemisty. Results: The positive rate of serum P53 Abs from the normal subjects was ≤2%. The positive rates of serum P53 Abs from the patients with tumors were obviously different according to the origin of tumor and its pathological stage. The positive rates of colorectal cacinoma, lung cancer, breast cancer, stomach cancer,primary liver cancer were 22%,34%,25%,36%,32%,respectively. Expression of P53 in 20 solid tumor specimens was assessed by immunohistochemistry and the positive rate was 60%.We also detected serum p53 Abs in 90 patients with colorectal cacinoma and breast cancer by Pharma Cell detecting kit.There was no significant difference between these 2 methods( P >0.05). Conclusion:This study shows that the positive rate of P53 Abs expression in solid tumor is significant higher than that in normal adult. It may become a new diagnostic indicator of tumor.