克隆酶均相免疫技术测定胃癌患者血清和粪便P53蛋白的研究

目的评价检测P53蛋白的克隆酶均相免疫分析方法的可行性。方法通过基因重组技术构建β半乳糖苷酶亚单位EA、ED及EDP53融合蛋白的原核表达质粒,建立克隆酶均相免疫技术,测定胃癌、非胃癌和正常对照组血清、粪便P53蛋白含量,并与酶联免疫吸附试验(ELISA)比较。结果31例胃癌患者的血清、粪便P53蛋白阳性率高于39例非胃癌患者和17名健康献血员(均P<001)。11例胃癌患者血清P53和免疫组化均阳性,且免疫复合物均以P53IgG形式存在,对5例胃癌血清动态观察表明,术后1周P53蛋白仍阳性,术后1个月显著下降,并在术后3个月消失。检测10份不同浓度的样本,表明克隆酶均相免疫法与ELISA具有较好的相关性。结论建立的克隆酶均相免疫技术可用于P53蛋白的测定,P53蛋白可作为判断胃癌预后的一项辅助诊断指标。

Establishment of cloned homogeneous immunoassay for detection of P53 protein in serum and stool samples from patients with gastric carcinoma

Objective To evaluate reliability of the cloned homogeneous immunoassay technique for detection of P53 protein content in serum and stool from patients with gastric carcinoma. Methods To prepare EA, ED and ED P53 fusion protein of beta galactosidase by PGEX 4T 2 expression vector, a novel cloned homegeneous immunoassay was establishment for detection of P53 protein in serum and stool samples from patients with gastric carcinoma and non gastric carcinoma groups as well as healthy control. Results The concentration of P53 protein in 31 gastric carcinoma subjects were remarkably increased as compared with 56 non gastric carcinoma ( P <0 01). Further studies showed that all cases were in accordance with immunohistochemical staining. Moreover, there existed P53 IgG immuno compound in these patients, these results implied that the entity type of P53 might contribute to prolong its halftime in peripheral circulation. There was preferential relation between cloned homegeneous immunoassay and ELISA in 10 samples. Conclusion Serum and stool P53 protein might be an independent biological marker in diagnosis of gastric carcinoma.