流式细胞术测12项细胞因子性能验证及评价

Verification and evaluation of the performance of 12-in-1 cytokine detection kit on flow cytometryne

目的对流式细胞术同时测12项细胞因子包括白细胞介素1β(IL-1β)、IL-2、IL-4、IL-5、IL-6、IL-8、IL-10、IL-12P70、IL-17、干扰素α(IFN-α)、IFN-γ、肿瘤坏死因子α(TNF-α)的性能进行验证及评价。方法参照WS/T492-2016《临床检验定量测定项目精度和正确度性能验证》、WS/T 402-2012《临床实验室检验项目参考区间的制定》,使用BD FACS CantoⅡ流式细胞仪对青岛瑞斯凯尔生物科技有限公司12项细胞因子检测试剂盒进行线性及范围、精密度、准确性、参考区间进行验证。结果IL-1β、IL-2、IL-4、IL-5、IL-6、IL-8、IL-10、IL-12P70、IL-17、IFN-α、IFN-γ、TNF-α在BD CantoⅡ流式细胞仪上检测结果在2.44~10000 pg/mL范围内呈线性。最低检测限浓度2.44 pg/mL下能良好检出,变异系数符合要求;批内精密度和批间精密度变异系数均≤15%;准确性验证相对偏差均在±15%以内,参考区间可直接使用。结论12细胞因子检测试剂盒(多重微球流式免疫荧光发光法)在BD CantoⅡ流式细胞仪的线性及范围、最低检测限、精密度、准确性及参考范围通过验证,可以满足临床检测需求。

Objective To verify and evaluate the performance of Flow cytometry used to detect 12 cytokines including IL-1β(IL-1β),IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12p70,IL-17,interferonα(IFN-α),IFN-γ,tumor necrosis factorα(TNF-α).Methods According to WS/T492-2016"Verification of Accuracy and Accuracy Performance of Clinical Test Quantitative Determination Project"and WS/T 402-2012"Formulation of Reference Section of Clinical Laboratory Test Project",BD FACS CantoⅡflow cytometer was used to verify the linearity,range,precision,accuracy,and reference interval of 12-in-1 cytokine detection kit of Qingdao Ruisikeer Biotechnology Co.,Ltd.Results IL-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12P70,IL-17,IFN-α,IFN-γ,TNF-αin the detection result on the BD CantoⅡflow cytometer was linear within the range of 2.44-10000 pg/mL.The minimum detection limit concentration can be well detected at a concentration of 2.44 pg/mL,and the coefficient of variation meets the requirements.Then,the coefficient of variation of intra-batch precision and inter-batch precision is≤15%.Besides,the relative deviation of accuracy verification is within±15%.So the reference interval could be used directly.Conclusion The 12-in-1 cytokine detection kit(multiple microsphere flow immunofluorescence method)has passed the verification of the linearity and range,minimum detection limit,precision,accuracy and reference range of the BD CantoⅡflow cytometer,which could satisfied the clinical detection demand.