毛细管电泳法在β地中海贫血患者 HbA1c和 HbA2检测中的应用

The application of capillary electrophoresis in HbA1c and HbA2 detection in beta thalassemia patients

目的探讨β地中海贫血对毛细管电泳法等8种糖化血红蛋白检测系统的影响，以及毛细管电泳糖化血红蛋白系统检测β地中海贫血中HbA2的应用。方法收集2014年1至8月北京大学深圳医院确诊的非β地中海贫血的样本338例，其中正常样本258名，α地中海贫血80例（基因型为－－SEA／αα）；以及β地中海贫血且不合并其他血红蛋白病样本（α珠蛋白基因正常，β珠蛋白基因异常且为杂合子）225例。采用毛细管电泳法（ Capillary 2 Flex Piercing）、酶法（ Norudia N HbA1c ）、免疫透射比浊法（ one HbA1c FS）、化学发光免疫法（ ARCHITECT HbA1c Reagent Kit）、亲和层析高压液相色谱法（Ultra2）和离子交换高压液相色谱法（HA8160、Variant ⅡTurbo和Variant ⅡTurbo 2．0），对正常和β地中海贫血各76例样本进行HbA1c检测，比较不同检测系统HbA1c结果的相关性。毛细管电泳糖化血红蛋白检测仪和毛细管电泳血红蛋白检测仪分别检测338例非β地中海贫血样本和225例β地中海贫血样本的HbA2，通过ROC曲线计算毛细管电泳糖化血红蛋白检测仪HbA2，建立筛查β地中海贫血的截断值。结果对于正常样本，7种检测系统与参比系统（ Ultra2）的相关性良好，相关系数都＞0．98；对于β地中海贫血样本，除Variant ⅡTurbo与参比系统相关系数为0．972外，其余检测系统相关系数都＞0．98。除Variant ⅡTurbo外，正常样本和β地中海贫血样本HbA1c结果的偏倚差异无统计学意义。毛细管电泳糖化血红蛋白检测系统和毛细管电泳血红蛋白检测系统的HbA2有良好的相关性，相关系数为0．993。毛细管电泳糖化血红蛋白检测系统HbA2筛查β地中海贫血的截断值设定为3．38时，敏感性为98．2％，特异性为100％。结论除Variant ⅡTurbo外，β地中海贫血对于其他糖化血红蛋白检测系统无明显影响。毛细管电泳糖化血红蛋白检测系统在检测糖化血红蛋白的同时，具有筛查β地中海贫血的功能。（中华检验医学杂志，2015，38：329－332）

Objective To investigate effects of β-thalassemia on 8 HbA1c measurement systems including Capillary electrophoresis and application of HbA2 values of samples with β-thalassemia from Capillary 2 Flex Piercing HbA1c measurement systems.Methods 338 samples without β-thalassemia （ include 80 α-thalassemia with --SEA/ααgenotype） and 225 samples with β-thalassemia were collected in Peking University Shenzhen hospital from January 2014 to August 2014.Samples withβ-thalassemia are all heterozygotes with normal α-globin and abnormal β-globin, and without combining other haemoglobinopathies.76 normal samples and 76 samples with β-thalassemia were selected from above all samples, HbA1c values of which were given by capillary electrophoresis （ Capillary Flex 2 Piercing）, enzymatic （ Norudia N HbA1c ）, immune transmission turbidity （ oneHbA1c FS ）, chemiluminescent immunoassay （ARCHITECT HbA1c Reagent Kit）, boronate affinity HPLC （Trinity Ultra2）and ion exchange HPLC （HA8160, VariantⅡTurbo and Variant Ⅱ BioRad 2.0）, respectively.HbA1c values given by the eight systems were compared.338 samples without β-thalassemia and 225 samples with β-thalassemia were measured by using Capillarys 2 Flex Piercing HbA1c system and capillary electrophoresis hemoglobin system, the cut-off values of HbA2 from Capillary 2 Flex Piercing HbA1c measurement system for β-thalassemia was 〈br〉 assessed by using ROC curve.Data statistics were done by using SPSS 19.0.Results The HbA1c values of normal samples were well correlated with the comparative system.The correlation coefficients for the 7 systems are all greater than 0.98.For samples withβ-thalassemia, the correlation coefficients are all greater than 0.98 except for Variant Ⅱ Turbo.The bias between normal samples and samples with β-thalassemia have no significant difference except for VariantⅡTurbo.HbA2 values from Capillary 2 Flex Piercing HbA1c measurement and capillary electrophoresis hemoglobin system were well correlated. The correlation coefficient is 0.993.When cut-off value of HbA2 from Capillary 2 Flex Piercing HbA1c measurement system for β-thalassemia is 3.38, sensitivity is 98.3%, specificity is 100% .Conclusions Samples withβ-thalassemia have no significant effects on HbA1c measurement syetems except for Variant Ⅱ Turbo.The Capillary 2 Flex Piercing HbA1c measurement syetem has the advantage of screeningβthalassemia by HbA2 while measuring HbA1c.