流式细胞仪检测结缔组织生长因子的方法学评价

摘要

目的:评价FACSArray流式细胞仪检测血清结缔组织生长因子(CTGF)的方法学性能.方法:用流式细胞仪测定血清高低浓度CTGF,计算两种浓度的批内变异系数、批间变异系数及相对偏差评价仪器的精密度、准确度及分析灵敏度,采用美国临床和实验室标准协会(CLSL)的有关要求进行评价;计算在校准品中加入聚苯乙烯微球、高浓度的甘油三酯(17 mmoL/L)、血红蛋白(12 g/L)、胆红素(170 umoL/L)或低浓度的甘油三酯(0.76 mmol/L)、血红蛋白(0.5 g/L)和胆红素(1.7 umol/L)干扰物对CTGF测定时的干扰率,对仪器分析性能进行验证和评价.结果:CTGF的高低两个浓度的批内变异系数分别为2.96％和3.75％,高低两个浓度的批间变异系数分别为9.02％和8.78％,CTGF的高低两个浓度的相对偏差分别为2.23％和1.85％,检测灵敏度为14.91 ng/L,均达到CLSL的有关要求;聚苯乙烯微球,高浓度的甘油三酯、血红蛋白及胆红素十扰物对CTGF测定时的干扰率分别为3.98％、12.76％、11.35％及11.14％,对检测有一定的干扰;低浓度的甘油三酯、血红蛋白及胆红素干扰物对CTGF测定时的干扰率分别为4.45％、3.49％和2.94％,对检测干扰相对于高值干扰物较小.结论:FACSArray流式细胞仪检测系统检测血清CTGF的性能指标可满足实验室认可的需求;高浓度的甘油三酯、血红蛋白和胆红素对CTGF的测定有一定干扰作用.%Objective:To evaluate methodology function of FACSArray flow cytometry test on the performance of connective tissue growth factor (CTGF).Methods:FACSArray flow cytometry system was used to detect the serum high and low concentration CTGF,calculate both concentration within assay variation,between assay variation,as well as precision,accuracy,analysis sensitivity of relative deviation evaluation equipment;adopting relevant requirements of CLSL for evaluation;after adding interference test to calibrator:polystyrene microspheres,high concentration triglyceride(17 mmol/L),hemoglobin (12 g/L),bilirubin (170 μ mol/L),or low concentration triglyceride (0.76 mmol/L),hemoglobin(0.5 g/L) and bilirubin(1.7 μmol/L),calculating disruption rate on CTGF detection and evaluating on the equipment analyzing performance.Results:The within assay variations of CTGF in both high and low concentration were 2.96％ and 3.75％ respectively.High and low concentrations between assay variations of CTGF were 9.02％ and 8.78％ respectively.The relative deviations of the high and low concentrations of CTGF were 2.23％ and 1.85％ respectively.The detection sensitivity of CTGF was 14.91 ng/L;all met the requirement of CLSL.Rate of interference in determination of CTGF of polystyrene microspheres,the high concentration of triglyceride,hemoglobin and bilirubin disrupters were 3.98％,12.76％,11.35％,and 1 1.14％ respectively,which showed certain interference on detection.Rate of interference in determination of CTGF of low concentration of triglyceride hemoglobin and bilirubin disrupters were 4.45％,3.49％ and 2.94％ respectively,which showed relatively minor interference on detection than high value disrupters.Conclusion:Performance index from FACSArray flow cytometry detecting serum CTGF could meet the requirements of laboratory.High concentration of triglyceride,hemoglobin and bilirubin had certain interference effect on the determination of CTGF.