College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China.
Affiliated Xiaoshan Hospital, Hangzhou Normal University, Hangzhou, China.
J Fluoresc. 2022 Jul;32(4):1501-1507. doi: 10.1007/s10895-022-02950-1. Epub 2022 May 5.
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 Eu 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.
建立基于时间分辨荧光免疫分析（TRFIA）的肿瘤相关胰蛋白酶原-2（TAT-2）快速、高敏感检测方法，并评估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，使用两种 TAT-2 抗体（包被抗体和 Eu 标记抗体）。然后建立、评价和应用 TAT-2-TRFIA 方法检测健康受试者和肺癌患者的血清 TAT-2 水平。TAT-2-TRFIA 方法的线性范围为 1.53-300ng/mL，批内变异系数（CV）在 1.67%-8.42%之间，批间 CV 在 4.29%-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 的交叉反应率分别为 0.02%和 0.82%。肺癌患者的血清 TAT-2 水平高于健康受试者（P<0.001）。与 TAT-2 联合后，CEA 和 CA-125 对肺癌的敏感性和特异性显著提高。结论：我们成功建立了一种高敏感的 TAT-2-TRFIA 方法，能够促进肺癌的及时诊断。
