TINIA检测HbA1c的性能评价及和IE-HPLC在HbE患者中的应用比较

摘要

目的：评价免疫抑制比浊法（TINIA）检测糖化血红蛋白（HbA1c ）的分析性能，与离子交换高效液相色谱法（IE-HPLC）检测HbA1c进行比对和偏倚评估，并评估血红蛋白（Hb）变异体E（HbE）对TINIA和IE-HPLC的影响。方法参考美国临床与实验室标准化协会（CLSI）发布的方法学评价系列文件（EP文件）及有关文献，对TINIA检测HbA1c的精密度、正确度、分析测量范围、分析干扰以及生物参考区间进行评价和验证，对TINIA与IE-HPLC检测Hb结构正常的血液样本和含HbE的血液样本进行对比分析和偏倚评估，分析患者平均血糖的相关性。结果 TINIA检测HbA1c的批内精密度＜2．08％，总不精密度＜2．94％；在HbA1c浓度为4．4％～18．3％范围内，理论值与实测均值呈线性相关（r＝0．9994）；Hb、游离胆红素（FBiL）、结合胆红素（CBiL）、乳糜（CH）浓度分别为9．9～49．6μmol/L、65．6～328．0μmol/L、65．6～328．0μmol/L和3000～12000 FTU时，差异百分比均＜5％；厂商提供的参考区间（4．8％～5．9％）适用于临床实验室。对于Hb结构正常的血液样本，TINIA和IE-HPLC的HbA1c检测结果差异无统计学意义（P＞0．05），两种方法的相关性良好（r2＝0．9992）。对于HbE浓度在5．4％～54．7％范围内的血液样本，TINIA的平均偏倚为-4．1％～1．9％，均＜5％，几乎不受HbE 的干扰， HbA1c检测结果与平均血糖的相关性较好（r＝0．9988）；当HbE＞6．4％时IE-HPLC检测HbA1c的实测值与理论值平均偏倚为5．7％～278．7％，差异百分比均＞5％，测定结果与预期理论结果有明显差异；两种方法检测结果差异有统计学意义（P＜0．01）。结论 TINIA检测HbA1c的分析性能符合临床测定的性能要求，不受HbE的干扰。当HbE＞6．4％时，IE-HPLC法检测HbA1c会受干扰，出现假性增高的情况。%Objective To evaluate the analysis performance of glycosylated hemoglobin A1c (HbA1c )detection by turbidimetric inhibition immunoassay (TINIA),to compare and evaluate the bias with ion exchange high performance liquid chromatography (IE-HPLC)for detecting HbA1c ,and to evaluate the influence of hemoglobin (Hb)E (HbE)on TINIA and IE-HPLC.Methods According to the methodology evaluation document (EP document)and relevant literatures from the Clinical and Laboratory Standards Institute (CLSI ),the precision,accuracy,detection range, analysis interference and biological reference interval analysis of HbA1c detection by TINIA were evaluated and verified. The comparison analysis and bias evaluation of TINIA and IE-HPLC for blood samples with normal Hb structure and blood samples in HbE patients were performed.The correlation with average blood glucose was analyzed.Results The within-run precision by TINIA was <2.08%,the total imprecision was <2.94%,and the theoretical value with detection mean value was linearly correlated in the concentration range of 4.4%-18.3%(r=0.9994).When Hb,free bilirubin(FBiL),conjugated bilirubin (CBiL)and chyle (CH)concentrations were 9.9-49.6 μmol/L,65.6-328.0 μmol/L,65.6-328.0 μmol/L and 3 000-12 000 FTU,the difference percentage was <5%.The manufacturers provided reference interval (4.8%-5.9%)being suitable for clinical laboratories.For the blood samples with normal Hb structure,there was no statistical significance for HbA1c detection between the 2 methods (P>0.05 ),and the results of HbA1c detection by TINIA and IE-HPLC had good correlation (r2 =0.999 2).For the blood samples with HbE concentration in the range of 5.4%-54.7%,the average bias by TINIA was -4.1%-1.9%,which was <5%.The interference was almost not affected by HbE,and the correlation of HbA1c detection results with average blood glucose was good (r=0.998 8).While HbE >6.4%,the average bias of the detection value and the theoretical value for HbA1c by IE-HPLC was 5.7%-278.7%,the difference percentage was >5%,and the expected theoretical results with the detection value had significant differences,and there was statistical significance between the 2 methods (P <0.01).Conclusions The performance of HbA1c detection by TINIA meets the performance requirements of clinical determination,and has a good correlation with that by IE-HPLC without interference by HbE.When HbE>6.4%,IE-HPLC for the detection of HbA1c has interference,and there will be false increasing.