诊断学理论与实践

诊断学理论与实践 ›› 2021, Vol. 20 ›› Issue (03): 271-278.doi: 10.16150/j.1671-2870.2021.03.008

• 论著 • 上一篇    下一篇

高效液相色谱技术联合红细胞参数在血红蛋白病筛查中的应用

王也飞1, 吴蓓颖2, 夏文权1, 陈宁1, 胡翊群3   

  1. 1.上海交通大学医学院附属瑞金医院检验系,上海 200025
    2.上海交通大学医学院附属瑞金医院检验科,上海 200025
    3.上海交通大学医学院,上海 200025
  • 收稿日期:2020-07-01 出版日期:2021-06-25 发布日期:2022-06-28

Application of high performance liquid chromatography (HPLC) and erythrocyte parameters in the screening of hemoglobinopathy

WANG Yefei1, WU Beiying2, XIA Wenquan1, HU Yiqun1   

  1. 1. Faculty of Medical Laboratory Science, Shanghai Jiao Tong University School of Medicine
    2. Department of Clinical Laboratory, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
    3. Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Received:2020-07-01 Online:2021-06-25 Published:2022-06-28

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摘要：

目的:探讨高效液相色谱（high performance liquid chromatography, HPLC）技术和红细胞参数在血红蛋白（hemoglobin, Hb）病筛查中的实际应用价值。方法:应用HPLC技术对1 029例临床疑似Hb病患者及进行溶血性贫血筛查者的血样本进行Hb分析,同时采用缺口聚合酶链反应、反向斑点杂交和珠蛋白基因DNA测序对样本进行基因分析;记录并分析患者的血常规红细胞参数。结果:1 029例血样本中,有591例（57.43%）确诊为珠蛋白生成障碍性贫血,14例（1.36%）确诊为异常Hb病。以基因分析结果为诊断金标准,应用HPLC检测β-珠蛋白生成障碍性贫血,以血红蛋白A2（hemoglobin A2, HbA2）3.9%为临界值, HbA2>3.9%诊断β-珠蛋白生成障碍性贫血的灵敏度为95.84%,特异度为96.87%,阳性预测值为95.40%,阴性预测值为97.20%,受试者操作特征曲线下面积（area under curve, AUC）为0.962;以HbA2 2.1%为临界值,HbA2≤2.1%结合出现快速区带来诊断中间型α-珠蛋白生成障碍性贫血[又称血红蛋白H（hemoglobin H, HbH）病]的灵敏度为100.00%,特异度为96.32%,阳性预测值为52.60%,阴性预测值为100.00%,AUC为0.990;以HbA2 3.2%为临界值,HbA2≤3.2%诊断静止型和标准型α-珠蛋白生成障碍性贫血的灵敏度为97.76%,特异度为53.11%,阳性预测值为24.0%,阴性预测值为99.4%,AUC为0.753。分别以平均红细胞体积（mean corpuscular volume, MCV）≤73.3 fl,平均红细胞血红蛋白量（mean corpuscular hemoglobin, MCH）≤23.5 pg,平均红细胞血红蛋白浓度（mean corpuscular hemoglobin contentration, MCHC）≤324 g/L诊断珠蛋白生成障碍性贫血,其灵敏度和特异度分别为87.98%和77.51%,93.73%和69.86%,89.20%和47.61%。结论:在红细胞参数测定的基础上,应用HPLC检测筛查β-珠蛋白生成障碍性贫血、HbH病和异常Hb病的灵敏度、特异度高,与基因分析结果间有较高的一致性,且操作简便、快速,适用于临床快速筛查;但对于静止型和标准型α-珠蛋白生成障碍性贫血,HPLC检测的筛查特异度和阳性预测值均较低,易造成漏诊。

关键词: 血红蛋白病, 高效液相色谱技术, 疾病筛查

Abstract:

Objective: To explore the application value of high performance liquid chromatography (HPLC) and erythrocyte parameters for screening of thalassemia in Shanghai. Methods: The hemoglobin of 1 029 samples from patients with suspected thalassemia and with suspected hemolytic anemia were analyzed using HPLC. Genetic analysis of the globin were performed simultaneously with GAP-PCR,reverse dot blot(RDB) and DNA sequencing. Results: Among 1029 samples, 591（57.43%） were diagnosed as having thalassemia and 14 (1.36%) having structural hemoglobin variants.Compared with results of genetic analysis,the sensitivity,specificity,positive predictive value and negative predictive value of HbA2 >3.9% by HPLC as cut-off value for diagnosing β-thalassemia were 95.84%, 96.87%, 95.40%, 97.20%,with AUC of 0.962. When HbA2≤2.1% by HPLC as the cutoff value combined with presence of rapid band were taken for diagnosing α-thalassemia moderate（HbH disease）, sensitivity, specificity, positive predictive value, negative predictive value were 96.32%, 52.60%, 100.00 %, with AUC of 0.990. If f HbA2 ≤3.2% by HPLC was taken as the cutoff value for diagnosing minor α-thalassemia or α-thalassemia trait, the corresponding indice were 97.76%, 53.11%, 24.0%, 99.4%,and AUC was 0.753. If MCV≤73.3 fl,MCH≤23.5 pg,MCHC≤324g/L were taken as the cutoff value alone for diagnosing thalassemia, sensitivity and specificity were 87.98%,77.51%, 93.73% and 69.86%, 89.20%,47.61%,respectively. Conclusions: Based on RBC parameters analysis,results of HPLC technology have a good consistence with the resluts of gene detection in the diagnosis of β-thalassemia, HbH disease and structural hemoglobin variant, while a low specificity and positive predictive value may occur in the diagnosis of minor α-thalassemia orα-thalassemia trait.

Key words: Hemoglobinopathy, High performance liquid chromatography, Disease screening

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