Bao Han, Fang Wei, Kao Shangbin, Wu Bin, Zheng Tianyu, Qin Yuan, Zhou Xiumei, Zhang Nenghua, Huang Biao
College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China.
Division of Laboratory Medicine, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Southern Medical University, Guangzhou, China.
J Fluoresc. 2025 Aug 18. doi: 10.1007/s10895-025-04507-4.
To develop a highly sensitive, wide-linear-range time-resolved fluorescence immunoassay (TRFIA) to detect serum YKL-40 and to measure its concentrations in patients with diabetic kidney disease (DKD). A double-antibody sandwich method was employed to establish the YKL-40 TRFIA, which utilizes two antibodies targeting distinct epitopes on the YKL-40 antigen (a coating antibody and a labeled antibody). The assay's linearity, sensitivity, accuracy, specificity, and recovery rate were evaluated. Serum YKL-40 concentrations were measured in healthy individuals and patients with DKD using this TRFIA. The YKL-40 TRFIA exhibited a linear range of 0.5-2000 ng/mL. Intra-assay and inter-assay coefficients of variation were 3.54-9.71% and 4.79-11.27%, respectively. The recovery rate was 101.73-102.62%. The assay showed no cross-reactivity with YKL-39, galectin-3 or CXCL9 and no serum interference. The YKL-40 TRFIA results were highly consistent with those of ELISA (P < 0.0001). The serum YKL-40 level in DKD patients was 314.78 (184.12, 482.49) ng/mL, significantly higher than that in healthy controls [65.73 (48.71, 98.70) ng/mL; P < 0.001]. This study successfully developed a highly sensitive and wide-linear-range method for YKL-40 detection, which can be used for clinical detection.
开发一种高灵敏度、宽线性范围的时间分辨荧光免疫分析法（TRFIA）以检测血清YKL-40，并测定糖尿病肾病（DKD）患者血清YKL-40的浓度。采用双抗体夹心方法建立YKL-40 TRFIA，该方法利用两种靶向YKL-40抗原不同表位的抗体（包被抗体和标记抗体）。对该检测方法的线性、灵敏度、准确性、特异性和回收率进行了评估。使用该TRFIA检测健康个体和DKD患者血清YKL-40浓度。YKL-40 TRFIA的线性范围为0.5-2000 ng/mL。批内和批间变异系数分别为3.54-9.71%和4.79-11.27%。回收率为101.73-102.62%。该检测方法与YKL-39、半乳糖凝集素-3或CXCL9无交叉反应，且无血清干扰。YKL-40 TRFIA结果与ELISA结果高度一致（P < 0.0001）。DKD患者血清YKL-40水平为314.78（184.12，482.49）ng/mL，显著高于健康对照者[65.73（48.71，98.70）ng/mL；P < 0.001]。本研究成功开发了一种高灵敏度、宽线性范围的YKL-40检测方法，可用于临床检测。
