College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China.
Wuxi No. 5 People's Hospital, Wuxi, China.
J Clin Lab Anal. 2021 May;35(5):e23758. doi: 10.1002/jcla.23758. Epub 2021 Mar 15.
To establish a time-resolved fluorescence immunoassay of interleukin (IL)-18 (IL-18-TRFIA) and detect its concentration in different liver disease serum samples.
The IL-18 coating antibody and the Eu -labeled detection antibody were used for the IL-18-TRFIA to detect serum IL-18 concentration in patients with liver cancer, hepatitis B, hepatitis C, autoimmune hepatitis, fatty liver disease, and healthy controls. The double-antibody sandwich method was used and methodological evaluation was performed.
The average intra- and inter-assay coefficient of variation for IL-18-TRFIA was 4.80% and 5.90%, respectively. The average recovery rate was 106.19 ± 3.44%. The sensitivity (10.96 pg/mL) was higher than that obtained using the ELISA method (62.5 pg/mL). The detection range was 10.96-1000 pg/mL. IL-6 and galectin-3 did not cross-react with IL-18-TRFIA. The serum concentration of IL-18 was (776.99; 653.48-952.39 pg/mL) in hepatitis C, (911; 775.55-1130.03 pg/mL) in fatty liver, (1048.88; 730.04-1185.10 pg/mL) in liver cancer, and (949.12; 723.70-1160.28 pg/mL) in hepatitis B. Moreover, IL-18 serum levels were significantly higher in patients than the healthy controls (483.09; 402.52-599.70/mL) (p < 0.0001). Autoimmune hepatitis with a serum IL-18 concentration of 571.62; 502.47-730.31 pg/mL was not significantly different from the healthy controls (p > 0.05).
We established a highly sensitive IL-18-TRFIA method that successfully detected serum IL-18 concentrations in different liver diseases. Furthermore, IL-18 serum concentration was higher in patients with liver cancer, hepatitis C, hepatitis B, and fatty liver disease compared to healthy controls.
建立白细胞介素（IL）-18（IL-18-TRFIA）时间分辨荧光免疫分析方法，并检测不同肝病血清样本中的浓度。
采用 IL-18 包被抗体和 Eu 标记检测抗体，建立 IL-18-TRFIA 检测肝癌、乙型肝炎、丙型肝炎、自身免疫性肝炎、脂肪肝和健康对照者血清 IL-18 浓度。采用双抗体夹心法进行方法学评价。
IL-18-TRFIA 的批内和批间变异系数分别为 4.80%和 5.90%。平均回收率为 106.19±3.44%。灵敏度（10.96pg/mL）高于酶联免疫吸附法（62.5pg/mL）。检测范围为 10.96-1000pg/mL。IL-6 和半乳糖凝集素-3 与 IL-18-TRFIA 无交叉反应。丙型肝炎患者血清 IL-18 浓度为（776.99；653.48-952.39pg/mL），脂肪肝患者为（911；775.55-1130.03pg/mL），肝癌患者为（1048.88；730.04-1185.10pg/mL），乙型肝炎患者为（949.12；723.70-1160.28pg/mL）。此外，与健康对照组相比，患者的血清 IL-18 水平显著升高（483.09；402.52-599.70pg/mL）（p<0.0001）。血清 IL-18 浓度为 571.62pg/mL 的自身免疫性肝炎与健康对照组无显著差异（p>0.05）。
我们建立了一种高灵敏度的 IL-18-TRFIA 方法，成功检测了不同肝病患者血清中 IL-18 的浓度。此外，与健康对照组相比，肝癌、丙型肝炎、乙型肝炎和脂肪肝患者的血清 IL-18 浓度更高。
