College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China.
Jiangnan University Medicine Center, Wuxi, China.
Anal Biochem. 2023 Sep 15;677:115252. doi: 10.1016/j.ab.2023.115252. Epub 2023 Jul 23.
11-Dehydro-thromboxane B2 (11-dehydro-TXB2) is the final stable metabolite of thromboxane A2 (TXA2) and is involved in thrombus formation. Patients with membranous nephropathy (MN) are prone to thromboembolism events.
Time-resolved fluorescence immunoassay (TRFIA) for 11-dehydro-TXB2 was established by indirect competitive method. The coated 11-dehydro-TXB2-BSA conjugate was used to bind the 11-dehydro-TXB2 antibody competitively to the 11-dehydro-TXB2 antigen in the samples, followed by Eu-labeled goat anti-mouse IgG antibody, to detect 11-dehydro-TXB2. This study measured 11-dehydro-TXB2 concentrations in serum samples from healthy individuals and patients with MN.
The linear range of TRFIA was 16.38-2000 pg/mL, the sensitivity was 4.70 pg/mL, the average coefficients of variation from intra-assay and inter-assay were 3.50% and 4.95%, respectively, and the recovery was 99.38%. The serum level of 11-dehydro-TXB2 in patients with MN was significantly higher than that in healthy subjects (P < 0.05). The serum 11-dehydro-TXB2 concentration detected by TRFIA was highly consistent with that by ELISA (ρ = 0.900).
This study successfully established a new highly sensitive method for the detection of 11-dehydro-TXB2 in serum. 11-Dehydro-TXB2 has great potential in evaluating the risk of thromboembolic events in patients with MN and is expected to be applied to other thromboembolic-related diseases.
11-脱氢血栓烷 B2（11-dehydro-TXB2）是血栓烷 A2（TXA2）的最终稳定代谢产物，参与血栓形成。膜性肾病（MN）患者易发生血栓栓塞事件。
采用间接竞争法建立 11-脱氢血栓烷 B2 的时间分辨荧光免疫分析（TRFIA）。用包被的 11-脱氢-TXB2-BSA 缀合物与样品中的 11-脱氢-TXB2 抗原竞争结合 11-脱氢-TXB2 抗体，然后加入 Eu 标记的山羊抗小鼠 IgG 抗体，检测 11-脱氢-TXB2。本研究测量了健康个体和 MN 患者血清样本中的 11-脱氢-TXB2 浓度。
TRFIA 的线性范围为 16.38-2000 pg/mL，灵敏度为 4.70 pg/mL，批内和批间变异系数的平均值分别为 3.50%和 4.95%，回收率为 99.38%。MN 患者血清中 11-脱氢-TXB2 水平明显高于健康受试者（P<0.05）。TRFIA 检测的血清 11-脱氢-TXB2 浓度与 ELISA 高度一致（ρ=0.900）。
本研究成功建立了一种检测血清中 11-脱氢-TXB2 的新的高灵敏度方法。11-脱氢-TXB2 在评估 MN 患者血栓栓塞事件风险方面具有很大的潜力，有望应用于其他血栓栓塞相关疾病。
