毛细管等电聚焦电泳定量分析健康成人及地中海贫血携带者的HbA_2  

Use of capillary electrophoresis to determine hemoglobin A_2 in healthy adults and α- and β-thalassemia carriers

作　　者：华亮[1] 李坚[2] 刘忠英[1] 钟惠珠[2] 廖灿[2] 徐湘民[1] 

机构地区：[1]第一军医大学医学遗传学教研室,广州510515 [2]广州市妇婴医院

出　　处：《中华医学遗传学杂志》2003年第5期421-424,共4页Chinese Journal of Medical Genetics

基　　金：广东省科委和广东省卫生厅联合攻关重大课题基金( 9 9B0 6 70 4 G)

摘　　要：目的　对等电聚焦毛细管电泳 (capillary isoelectric focusing,CIEF)检测血红蛋白 A2 (HbA2 )含量做出用于筛查地中海贫血 (地贫 )携带者的方法学评价。方法　收集 10 5名健康成人、93例经常规地贫筛查有阳性表型的连续临床样品。通过正常人样品分析比较 CIEF测定 Hb A2 与 Helena Spife combo电泳系统结果的一致性 ;通过比较样品内和样品间 Hb A2 的 CIEF测定值的变异系数评价该法的精确度和稳定性 ;通过分析阳性样品的基因型来评价 CIEF检测地贫的准确性。结果　用 CIEF分析当地健康人的Hb A2 正常参考值为 3 .59%～ 5.2 3 %。CIEF结果与常规血红蛋白电泳结果具有良好的线性关系 ;以基因分析为确诊标准 ,用 CIEF检测 Hb A2 含量 ,及结合红细胞指数阳性测定值所筛查的 87例 α-地贫或 β-地贫携带者、6例成人血红蛋白 E病例样品均被准确诊断 ,未见假阳性或假阴性结果。结论　等电聚焦毛细管电泳是一种快速高效、高稳定性、高精确度的 Hb A2 测定方法 。Objective: To evaluate the capillary isoelectric focusing (CIEF) method for the estimation of blood hemoglobin A2 (Hb A2) concentrations in routine thalassemia screening. Methods: A total of 105 samples from healthy adults and 93 samples with positive phenotypes were collected by routine thalassemia screening. CIEF was compared with Helena spife combo electrophoresis system for Hb A2 measurement and its precision and reproducibility were tested by analyzing intra-assay or inter-assay coefficient of variations (CVs). The reliability and veracity of Hb A 2 measurement by CIEF for the detection of α- and β-thalassemia including Hb E were evaluated by genotyping of 93 consecutive samples for routine thalassemia screening. Results: By use of CIEF for measurement of Hb A2 in a local healthy adult population, the range of reference value(3. 59%-5.23%) was obtained. The results of CIEF showed good linearity relation to that of conventional Hb electrophoresis assay. All thalassemia carriers (43 cases of α-thals and 44 of β-thals) or Hb E carriers (6 cases) presumptively identified by the present CIEF for the quantification of Hb A2, combined with routine RBC parameters for indicating microcytosis and hypochromia were confirmed to be the heterozygous or compound heterozygous defects of α- or β-globin gene by molecular diagnosis, without any false positive or false negative results. Conclusion: The measurement of Hb A2 by CIEF method is rapid, precise and reproducible; it could be used in routine screening for α- or β- thalassemia.

关 键 词：等电聚焦毛细管电泳 地中海贫血 血红蛋白A2 CIEF HBA2 红细胞 

分 类 号：R556[医药卫生—血液循环系统疾病]