流式微球捕获芯片技术检测血浆细胞因子方法学性能验证

Methodological performance verification of flow microsphere capture chip technology for detecting plasma cytokines

目的对流式微球捕获芯片技术检测血浆中多种细胞因子方法学的性能进行验证。方法依据中华人民共和国卫生行业及美国临床和实验室标准协会的标准文件,应用流式微球捕获芯片技术检测血浆中的白细胞介素-1β(IL-1β)、IL-2R、IL-6、IL-8、IL-10和肿瘤坏死因子α(TNF-α)6种细胞因子,并对其精密度、准确度、线性范围、参考区间、最低检测限、抗干扰能力进行评价。结果6项细胞因子批内精密度变异系数(CV)为1.50%~8.83%,批间精密度CV为2.48%~6.75%,准确度相对偏差≤±15%,IL-1β、IL-6、IL-8、IL-10和TNF-α在2.5~5000.0 pg/mL的浓度范围内,IL-2R在5~7500 U/mL的浓度范围内,线性范围良好。生物参考区间验证结果显示,90%的结果在厂家提供的生物参考区间范围内。IL-1β、IL-6、IL-8、IL-10、TNF-α的最低检出限为≤2.5 pg/mL,IL-2R最低检出限为≤5 U/mL,干扰物质(甘油三酯10 mmol/L、血红蛋白5 mg/L和胆红素258μmol/L)对试剂盒检测结果无影响。结论在mindray Bricyte E6流式细胞仪上应用流式微球捕获芯片技术检测血浆细胞因子的方法性能较好,可为临床提供准确可靠的6种细胞因子的定量检测结果。

Objective To verify the performance of the flow microsphere capture chip technology in the detection of various cytokines in plasma.Methods According to the standard documents of the health industry of the People′s Republic of China and the American Society of Clinical and Laboratory Standards,the six cytokines of interleukin(IL)-1β,IL-2R,IL-6,IL-8,IL-10 and tumor necrosis factorα(TNF-α)in plasma were detected by flow microsphere capture chip technology,and the precision,accuracy,linear range,reference interval,minimum detection limit and anti-interference ability were evaluated.Results The intra-batch precision coefficient of variation(CV)of the six cytokines was 1.50%-8.83%,and the inter-batch precision(CV)was 2.48%-6.75%.The relative deviation of accuracy was≤±15%.The concentration range of IL-1β,IL-6,IL-8,IL-10 and TNF-αwas 2.5-5000.0 pg/mL,and the concentration range of IL-2R was 5-7500 U/mL.The linear range was good.The verification results of biological reference interval showed that 90%of the results were within the range of biological reference interval provided by the manufacturer.The minimum detection limits of IL-1β,IL-6,IL-8,IL-10 and TNF-αwere≤2.5 pg/mL,and the minimum detection limit of IL-2R was≤5 U/mL.The interfering substances(triglyceride 10 mmol/L,hemoglobin 5 mg/L and bilirubin 258μmol/L)had no effect on the test results of the kit.Conclusion The method of detecting plasma cytokines by flow cytometric bead capture chip technology on mindray Bricyte E6 flow cytometry has good performance,which can provide accurate and reliable quantitative detection results of six cytokines for clinical practice.