Hb E变异体对5种糖化血红蛋白检测系统测定结果的干扰评价

Interference of hemoglobin E on measurements of glycosylated hemoglobin （ HbAlc ）

目的评价血红蛋白变异体E（HbE）对5种糖化血红蛋白（HbA1e）检测系统测定结果的干扰。其中离子交换高效液相色谱（IE—HPLC）法2种、硼酸盐亲和层析高效液相色谱法（AC—HPLC）、免疫抑制比浊法（TINIA）和酶法（EM）各1种。方法观察性研究。收集2012年5月至2013年10月来自中山大学附属中山医院的60份全血样本，分为健康对照组（20份）、糖尿病组（20份）和HbE变异体组（20份），分别用5种检测系统检测全血HbA1e，浓度，依据美国国家糖化血红蛋白标准化计划（NGSP）的判定标准，对不同组别5种检测系统的检测结果进行比对分析和偏倚评估，对HbE变异体组各检测系统HbA1e。检测结果计算得出的估计平均血糖值（eAG）与空腹血糖（FPG）水平差异进行统计学分析，对检测结果进行Deming回归分析，确定HbE对HbA，。检测结果是否具有显著临床影响，将6％与9％HbA1e。±10％相对偏差作为评价范围。采用配对t检验进行统计学分析。结果正常对照组和糖尿病组4种检测系统与通过NGSPI级实验室认证的比较系统差值的95％置信区间（95％CI）差异均在比较系统的±0．7％HbA1e以内、偏差％均小于6％，测定结果差异无统计学意义（P〉0．05）。HbE变异体组AC—HPLC法、Variant11IE—HPLC法和TINIA法3种检测系统HbA1e结果计算得出的eAG和FPG水平差异无统计学意义（P〉0．05），而VariantⅡ TurboIE．HPLC和酶法2个检测系统结果计算得出的eAG水平与FPG水平差异均具有统计学意义（P〈0．01）。BioradVariant Ⅱ IE—HPLC法、TINIA法与比较系统AC—HPLC法测定结果差值的95％CI差异落在比较系统的±0．7％HbA1e。以内、偏差％分别为-2．0％～5．6％和-5．8％～4．0％，测定结果差异无统计学意义（P〉0．05），且6％与9％HbA。浓度时，检测方法及比较系统样本的平均差异均小于临床可接受范围，结果显示这2种方法不受HbE的干扰。而BioradVariant Ⅱ TurboIE—HPLC法和酶法2种检测系统与比较系统AC—HPLC法测定结果的95％凹差异均落在比较系统的±0．7％HbA1e以外、偏差％分别为13．9％～40．1％和-34．1％-1．3％，均大于6％，测定结果差异均具有显著统计学意义（P〈0．01）。BioradVariant Ⅱ TurborIE—HPLC法与比较系统相比呈正偏差，酶法与比较系统相比呈负偏差。且6％与9％HbA1e浓度时，检测方法及比较系统样本的平均差异均大于临床可接受范围，HbE对这2种方法具有显著临床干扰。结论HbE对不同HbA1e，检测系统的干扰程度不同，临床实验室在进行HbA1e。检测时，应注意识别Hb变异成分，选用合适的方法进行HbA1e。测定或替代指标以防止干扰的发牛。（中华检验医学杂杀．2014．37：921．927）

Objective To evaluate the interference of hemoglobin variant E （HbE） on five kinds of glycosylated hemoglobin （HbA1e） system. Two of them were ion-exchange high performance liquidchromatography （IE-HPLC） methods, one was affinity chromatography high performance liquid chromatography （AC-HPLC） method, one was turbidimetric inhibition immunoassay method （TINIA）, one was enzyme （EM） method. Methods All 60 blood samples from May 2012 to October 2013 were collected from Zhongshan Hospital of Yat-sen University, and then divided into normal control group （20 cases）, diabetic group （20 cases） and HbE group （20 cases）. Variants were used to detect the whole blood concentration of HbAtc by five detection systems. Based on the judgment standards of National Glycohemoglobin Standardization Program （NGSP）, comparison analysis and bias evaluation results for different groups of 5 kinds of detection systems were estimated. The statistical difference of mean blood glucose （eAG） and fasting blood glucose （FPG） was calculated using the estimation of HbE variant group detection system HbAlo results. The test results were analyzed by Deming regression analysis, to determine whether HbE has significant clinical effect on the results of HbAlc, using 6% HbA1e and 9% ± 10% relative error as the evaluation scope. Results The 95% confidence interval （95% CI） of the 4 kinds of detection system in normal control group and diabetic group were within ± 0.7% HbA1e deviation, which was less than 6%, comparing to NGSP I laboratory certification, indicating no significant difference of determination resuhs（ P 〉 0.05 ）. There were no significant difference between eAG and FPG levels in HbE group using AC-HPLC, Variant HbE variant Ⅱ IE-HPLC method and TINIA method（P 〉 0.05 ）. While the difference between eAG level and FPG level of Variant Ⅱ Turbo IE-HPLC and enzymatic method was statistically significant （P 〈 0.01 ）. Bio-rad VariantⅡ IE-HPLC method, TINIA method and AC-HPLC method for the determination of the difference of control system 95% CI difference fall in the control system of ±0.7% HbA1e, and deviations were - 2.0% - 7.0% and - 7.1% - 4.0%, indicating no significant difference of determination results （ P 〉 0. 05 ）. Also when in the 6% and 9% HbA1e, concentrations, detection method and average differences between control samples were less than the clinically acceptable range,indicating that the interference of these 2 methods was not affected by HbE. The difference of 95% CIof Bio-Rad Variant Ⅱ Turbo IE-HPLC method and enzyme method were located outside the ± O. 7% HbA1e, comparing to AC-HPLC method. The determinations of the deviation were 13.9% ±40. 1% and -34.1%±1.3% respectively, which were greater than 6%. The difference was of statistically significance（P 〈 0.01 ）. Comparing to the control system, Bio-rad Variant 1I Turbor IE-HPLC held the positive deviation, while the enzyme method held the negative deviation. Also when in the 6% and 9% HbA1e concentrations, the mean differences of samples under detection method and control method were both greater than the clinically acceptable range, indicating that HbE has significant clinical interference in these 2 kinds of methods. Conclusion HbE had different interference effects in clinical laboratory for HbAI,, test, so one should pay attention to this kind of Hb variance in daily practice. Appropriate method should be selected to prevent the occurrence of interference to the determination or to find a surrogate marker of HbA1e. （Chin J Lab Med,2014,37：921-927）