对用于检测血红蛋白变体以及定量测定血红蛋白A、A2、E和F的超高效液相色谱法（PR-HPLC）与毛细管区带电泳法（CZE）分析性能的比较评估。

A comparative evaluation of the analytical performances of premier resolution-high-performance liquid chromatography (PR-HPLC) with capillary zone electrophoresis (CZE) assays for the detection of hemoglobin variants and the quantitation of HbA, A, E, and F.

作者信息

Laksap Sirikwan, Suanboon Suphisara, Punyamung Manoo, Ruengdit Chedtapak, Pornprasert Sakorn

机构信息

Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.

Associated Medical Sciences Clinical Service Center, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.

出版信息

Clin Chem Lab Med. 2024 Jan 11;62(7):1383-1392. doi: 10.1515/cclm-2023-1458. Print 2024 Jun 25.

DOI:10.1515/cclm-2023-1458

PMID:38197862

Abstract

OBJECTIVES

Hemoglobinopathies, including thalassemia and hemoglobin (Hb) variants, are common hematological disorders in tropical countries. Accurate and precise separation of hemoglobin types and reliable quantitation are necessary for differential diagnosis of these disorders.

METHODS

We have evaluated the analytical performances of premier resolution-high-performance liquid chromatography (PR-HPLC; Trinity Biotech, Co. Wicklow, Ireland) to assist in the presumptive diagnosis of thalassemia and Hb variants commonly found in Southeast Asian countries. HbA, HbA, HbE, and HbF levels were separated and quantified in 120 blood samples from unrelated adult subjects and compared with those analyzed by capillary zone electrophoresis (CZE; CAPILLARYS™ 2, Sebia, Norcross, GA, US). The Hb analysis patterns of Hb variants obtained from the PR-HPLC system were also compared to those obtained from HPLC (VARIANT II, β-thalassemia Short Program, Bio-Rad, Laboratories, Hercules, CA, US) and CZE systems.

RESULTS

The PR-HPLC had excellent precision with a coefficient of variation (CV) for HbA quantitation of 3.8 % within-run and 5.2 % between-run. The levels of HbA/E quantified by the PR-HPLC system correlated well with those of the CZE system (=0.997). In addition, thalassemia interpretation results obtained from the PR-HPLC and the CZE showed 100 % agreement. Moreover, chromatograms of the PR-HPLC were also comparable to those of VII-HPLC and CAP2-CZE electropherograms.

CONCLUSIONS

The PR-HPLC system would be applicable to diagnose common forms of thalassemia and Hb variants in Southeast Asia.

摘要

目的

血红蛋白病，包括地中海贫血和血红蛋白（Hb）变异体，是热带国家常见的血液系统疾病。准确、精确地分离血红蛋白类型并进行可靠的定量分析对于这些疾病的鉴别诊断至关重要。

方法

我们评估了高级分辨率高效液相色谱法（PR-HPLC；Trinity Biotech公司，爱尔兰威克洛）的分析性能，以协助对东南亚国家常见的地中海贫血和Hb变异体进行初步诊断。对120例无关成年受试者的血液样本中的HbA、HbA₂、HbE和HbF水平进行分离和定量，并与毛细管区带电泳（CZE；CAPILLARYS™ 2，Sebia公司，美国佐治亚州诺克罗斯）分析的结果进行比较。还将PR-HPLC系统获得的Hb变异体的Hb分析模式与HPLC（VARIANT II，β-地中海贫血短程序，Bio-Rad公司，美国加利福尼亚州赫拉克勒斯）和CZE系统获得的模式进行了比较。

结果

PR-HPLC具有出色的精密度，HbA定量的批内变异系数（CV）为3.8%，批间变异系数为5.2%。PR-HPLC系统定量的HbA₂/E水平与CZE系统的水平相关性良好（r = 0.997）。此外，PR-HPLC和CZE获得的地中海贫血解释结果显示100%一致。此外，PR-HPLC的色谱图也与VII-HPLC和CAP2-CZE电泳图相当。