高效液相色谱法定量分析HbA_2在珠蛋白生成障碍性贫血筛查中的应用

Quantitative analysis of HbA_2 with high-performance liquid chromatography in thalassemia screening

目的 对高效液相色谱法（HPLC）定量分析血红蛋白A2（HbA2）用于筛查β珠蛋白生成障碍性贫血（地中海贫血,地贫）基因携带者做出方法学评价.方法 地贫筛查中随机选择的75对夫妇及140对地贫高危夫妇,以地贫基因分析结果为金标准,通过比较样品内和样品间HbA2的HPLC测定值的变异系数评价该法的精密度和稳定性;通过分析阳性样品的基因型评价HPLC检测地贫的准确性.结果 用HPLC分析当地健康成人的HbA2的95%频数分布范围为2.28%～3.42%,HPLC诊断β地贫杂合子的HbA2临界值（cut off value）为≥4.35%.在基因分析确诊的65例β地贫杂合子、19例αβ复合地贫、2例HbE病及8例HbH病中,HPLC仅1例漏诊,无误诊.结论 高效液相色谱法是一种快速、稳定性强、精密度好的HbA2测定方法,可用于β地贫杂合子的筛查.

Objecitve To evaluate the application of quantitative analysis of HbA2 with high-performance liquid chromatography （HPLC） in thalassemia screening. Methods Seventy-five couples randomly selected and 140 couples at high risk of thalassemia were studied. Gene analysis of thalassemia was adopted as＂ gold standard＂. The precision and stability were evaluated by comparing the coefficient of variation intra-assay and inter-assay of HbA2 value measured with HPLC. The genotypes of positive samples＂ were determined to assess the accuracy of HPLC-measured HbA2 for diagnosis of thalamessia. Results The distribution range of 95% frequency of HbA2 measured by HPLC in the local healthy adults was from 2.28% to 3.42%. The cutoff value of HbA2 for diagnosing heterozygous 13 thalassemia was greater than or equal to 4.35 %. Among 94 patients （65 cases of heterozygous 13 thalassemia, 19 cases of 13 thalassemia,2 cases of HbE and 8 cases of HbH） only 1 case was lost,and no one was misdiagnosed. Conclusion HPLC is a fast,highly stable and accurate method for measuring HbA2 so it can be adopted in screening 13 thalassemia.