非抗体结合蛋白-抗体组合的GPC3双夹心化学发光法在临床的初步应用

Clinical application of a non-antibody binding protein and antibody combined GPC3 sandwich chemiluminescence immunoassay

目的:探讨非抗体结合蛋白-抗体双夹心化学发光免疫诊断试剂检测磷脂酰肌醇蛋白聚糖-3(GPC3)在肝细胞癌(HCC)血清学的临床诊断价值和鉴别诊断价值.方法:应用本课题前期研制的非抗体结合蛋白-抗体GPC3夹心化学发光法诊断试剂检测HCC、肝内胆管癌、胃癌、肺癌、乳腺癌、正常人群等患者血清GPC3水平,同时用GPC3免疫组织化学(IHC)检测上述疾病患者的肿瘤组织,同时分析甲胎蛋白(AFP)和本实验方法对HCC的诊断效能,进一步分析GPC3表达于不同细胞结构部位与血清水平关系.结果:非抗体结合蛋白-抗体GPC3双夹心化学发光试剂盒检测HCC组GPC3血清水平明显高于其他组(>21倍,P值均小于0.01),与GPC3 IHC结果相一致.联合GPC3和AFP的曲线下面积(AUC)为0.922,明显高于单一标志物(GPC3和AFP的AUC分别为0.899和0.821,P值均小于0.05)的诊断价值.GPC3主要表达于胞膜和混合表达(胞膜和胞质),且所有胞膜和混合表达阳性患者血清样本的GPC3均高水平表达.结论:非抗体结合蛋白-抗体GPC3双夹心化学发光试剂盒具有良好的诊断特异性及准确性,有可能用于HCC的鉴别诊断,且对分析GPC3胞膜型HCC的诊断、疗效评价及预后提供更多诊断价值信息.

Aim： To evaluate the glypican 3（GPC3） chemiluminescence immunoassay（CLIA） kit based on combination of a non-antibody binding protein and a monoclonal antibody（GPC3-CLIA） for diagnosis and differential diagnosis of Hepatocellular Carcinoma（HCC）. Methods： Human sera from healthy individuals and patients with HCC,intrahepatic cholangiocarcinoma,gastric cancer,lung canceror breast cancer were tested by GPC3-CLIA kit developed by our previous work,whereas tissue samples from above patients were also collected and detected by immunohistochemistry（IHC）. Moreover alpha fetal protein（AFP） was used as another tumor marker to assess the efficacy of our GPC3-CLIA kit.Finally,relevant analysis for specific expression location of GPC3 in the tumor tissues and its serum levels was conducted. Results： Serum GPC3 levels detected by GPC3 CLIA kit in HCC patients were significantly higher than other cancer patients and healthy individuals（P 0. 01）,which was consistent with IHC results. The area under curve（AUC） of the combination of GPC3 and AFP was significantly larger than that of GPC3 or AFP（0. 922 vs. 0. 899,and 0. 821,P 0. 05,respectively）. GPC3 was expressed mainly in the cytomembrane and the mixture（cytomembrane and cytoplasm）,which showed higher serum GPC3 levels. Conclusion： GPC3-CLIA kit is sensitive,specific and accurate for HCC diagnosis,which could be a good candidate method for HCC differential diagnosis,and it may be also useful for evaluating therapeutic efficacy and prognosis of HCC patients with GPC3 positive in the cell membrane.