高效液相色谱法检测糖化血红蛋白性能验证

Performance verification for high performance liquid chromatography detection of glycated hemoglobin

目的：对 BIO-RAD D-10高效液相色谱法检测糖化血红蛋白（HbA1c）的性能进行验证,评价其检测性能是否满足实验室质量及临床诊疗的要求。方法按 CLSI EP15A2文件的方法对糖化血红蛋白进行正确度、精密度的验证,采用线性回归分析方法对线性进行验证,按 WS/T 4022012的方法对参考区间进行验证。结果HbA1c 校准品水平1的标识值为5.3%,测定结果的验证限为1.63%-8.78%,校准品水平2的标识值为10.0%,测定结果的验证限为4.76%-13.64%,校准品的标识值均包含在测定值的95%置信区内,正确度验证通过。HbA1c 浓度为5.24%的实验室变异系数（CV）为1.01%,HbA1c 浓度为9.84%的 CV 为0.54%,均小于厂商声明的精密度,精密度验证通过。 HbA1c 线性校准品的实测值和目标值的线性回归方程为 Y =1.008X ＋0.023,相关系数指数 r 2为0.999,线性良好。20份健康成人标本的 HbA1c 检测结果均落在制造商提供的生物参考区间4.1%-6.2%范围内,参考区间验证通过。结论BIO-RAD D-10高效液相色谱法检测 HbA1c 的正确度、精密度、线性、参考区间均通过验证,能为临床提诊疗供准确可靠的检验结果,满足临床检测的要求,可用于临床标本的检测。

Objective To verify the detection performance of glycated hemoglobin （HbA1c）by using the BIO-RAD D-10 high performance liquid chromatography method,and to evaluate whether the detection performance can satisfy the requirements of the laboratory and clinical diagnosis and treatment.Methods The method of CLSI EP1 5A2 was used to verify the accuracy and precision.The linear correlation and reference range were verified respectively by linear regression analysis and the method of the WS/T 402 2012.Results The calibration value of HbA1c calibrator level 1 was 5.3% and the validation limit of measured results was 1. 63%-8.78%.The calibration value of HbA1c calibrator Level 2 was 10.0%,and the validation limit of measured results was 4.76%-13.64%.Both calibration values were included in the 95% confidence range of measured values,so the accuracy verification passed.Laboratory variation coefficient （CV）for HbA1c with concentration of 5.24% was 1.01% and for 9.84% was 0. 54%,both less than the precision that the manufacturer declared,so the precision verification passed.The equation of linear regression analysis for HbA1c linearity calibrator measured value and target value wasY =1.008X ＋0.023,with correlation coefficient index r 2 of 0.999,so the linearity was good.The measured results of 20 healthy adults specimens were in the reference interval（4.1%-6.2%）that the manufacturer provided.Conclusion Accuracy,precision,linear correlation and reference range of HbA1c measured by BIO-RAD D-10 HPLC detection method were verified.It can provide the accurate and credible measurement results for clinical diagnosis and treatment,and can be used in the detection of clinical specimens.