College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China.
Wuxi People's Hospital Affiliated to Nanjing Medical University, Wuxi, China.
J Clin Lab Anal. 2020 Jun;34(6):e23248. doi: 10.1002/jcla.23248. Epub 2020 Feb 20.
We aimed to develop a time-resolved fluorescence immunoassay (TRFIA) for detecting soluble T-cell immunoglobulin and mucin domain 3 (sTim3) in serum samples and to demonstrate a preliminary application of this method in membranous nephropathy (MN).
sTim3 TRFIA was developed, and the sTim3 concentration in the serum of patients with MN and healthy individuals was detected using a sandwich method.
The sensitivity of the developed sTim3 TRFIA was 0.66 ng/mL, higher than that of an enzyme-linked immunosorbent assay (ELISA) (1.11 ng/mL). The detection range was 0.66-40 ng/mL. The intra-assay coefficient of variation (CV) for sTim3 was 1.64%-4.68%, and the inter-assay CV was 5.72%-9.32%. The cross-reactivity to interleukin 6 (IL-6) and kidney injury molecule 1 (KIM-1) was 0.25% and 0.04%, respectively. The average recovery was 105.26%. The sTim3 concentration in patients with MN was considerably higher than that in healthy individuals (P < .001). The sTim3 concentration in the serum of patients with MN was significantly increased from G1 to G4 based on the Jonckheere-Terpstra test (P < .001). Thus, we used sTim3 as a diagnostic indicator for distinguishing between healthy individuals and patients with MN as well as between different stages of MN.
We successfully established TRFIA to detect sTim3 in serum. We then applied this method to patients with MN, demonstrating for the first time that TRFIA is a valid diagnostic tool to detect sTim3 in serum.
我们旨在开发一种用于检测血清样本中可溶性 T 细胞免疫球蛋白和粘蛋白结构域 3（sTim3）的时间分辨荧光免疫分析（TRFIA），并展示该方法在膜性肾病（MN）中的初步应用。
建立 sTim3 TRFIA，采用夹心法检测 MN 患者和健康个体血清中 sTim3 的浓度。
所开发的 sTim3 TRFIA 的灵敏度为 0.66ng/mL，高于酶联免疫吸附测定（ELISA）（1.11ng/mL）。检测范围为 0.66-40ng/mL。sTim3 的批内变异系数（CV）为 1.64%-4.68%，批间 CV 为 5.72%-9.32%。与白细胞介素 6（IL-6）和肾损伤分子 1（KIM-1）的交叉反应分别为 0.25%和 0.04%。平均回收率为 105.26%。MN 患者的 sTim3 浓度明显高于健康个体（P<0.001）。根据 Jonckheere-Terpstra 检验，MN 患者的 sTim3 浓度从 G1 到 G4 显著增加（P<0.001）。因此，我们使用 sTim3 作为区分健康个体和 MN 患者以及 MN 不同阶段的诊断指标。
我们成功建立了用于检测血清中 sTim3 的 TRFIA，并首次将该方法应用于 MN 患者，证明 TRFIA 是一种有效的血清 sTim3 诊断工具。
