Abstract：
To establish a rapid and highly sensitive assay for tumor-associated trypsinogen-2 (TAT-2) based on the time-resolved fluorescence immunoassay (TRFIA) and evaluate its potential clinical value in patients with lung cancer. The double-antibody sandwich method was used in detecting TAT-2 antigen concentrations, and two types of TAT-2 antibodies (coating antibodies and Eu3+ labeled antibodies) were used. A TAT-2-TRFIA method was then established, evaluated, and used in detecting the serum TAT-2 levels of healthy subjects and patients with lung cancer. The linear range of the TAT-2-TRFIA method was 1.53-300 ng/mL, the intra-assay coefficient of variation (CV) were between 1.67% and 8.42%, and the inter-assay CV were between 4.29% and 11.44%. The recovery rates of TAT-2-TRFIA were between 99.17% and 107.06%. The cross-reactivities of trypsin and T-cell immunoglobulin mucin 3 were 0.02% and 0.82%, respectively. The serum TAT-2 levels of patients with lung cancer were higher than those of healthy subjects (P < 0.001). Combined with TAT-2, the sensitivity and specificity of CEA and CA-125 for lung cancer improved significantly. Conclusion: We successfully established a highly sensitive TAT-2-TRFIA method, which was able to facilitate the timely diagnosis of lung cancer.
摘要：
建立基于时间分辨荧光免疫分析法(TRFIA)的肿瘤相关胰蛋白酶原-2(TAT-2)快速检测方法，评价其在肺癌患者中的潜在临床价值。采用双抗体夹心法检测TAT-2抗原浓度,并且使用两种类型的TAT-2抗体(包被抗体和Eu3+标记的抗体)。然后建立了TAT-2-TRFIA方法，评估，并用于检测健康受试者和肺癌患者的血清TAT-2水平。TAT-2-TRFIA法的线性范围为1.53-300ng/mL,测定内变异系数（CV）在1.67％至8.42％之间，测定间CV在4.29%和11.44%之间。TAT-2-TRFIA的回收率在99.17%～107.06%之间。胰蛋白酶和T细胞免疫球蛋白粘蛋白3的交叉反应性分别为0.02%和0.82%,分别。肺癌患者血清TAT-2水平高于健康者(P<0.001)。CEA和CA-125联合TAT-2对肺癌的敏感性和特异性明显提高。结论:我们成功建立了灵敏度高的TAT-2-TRFIA方法,能够促进肺癌的及时诊断。
