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Detection of coinherited Hb H-Constant Spring/Paksé disease and Hb E by capillary electrophoresis and high
Sakorn Pornprasert1, Jarurin Waneesorn
1Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang-Mai University, Chiang-Mai, Thailand. sakornmi001@gmail.com
Insights
Capillary electrophoresis (CE) excels over high-performance liquid chromatography (HPLC) in detecting Hb H-Constant Spring/Paksé-E disease. CE accurately quantifies Hb Bart's and Hb CS/Paksé, aiding diagnosis and prevention programs.
Area of Science:
- Clinical Chemistry and Hematology
- Analytical Biochemistry
- Molecular Diagnostics
Background:
- Hemoglobinopathies, including Hb H disease, require accurate diagnostic methods for effective management.
- Existing methods like HPLC have limitations in detecting and quantifying specific complex hemoglobin variants.
- Capillary electrophoresis (CE) offers potential advantages in hemoglobin analysis.
Purpose of the Study:
- To compare the efficacy of Capillary Electrophoresis (CE) and High-Performance Liquid Chromatography (HPLC) in detecting Hb H-Constant Spring/Paksé-E disease.
- To evaluate the quantification capabilities of CE and HPLC for specific hemoglobin variants.
- To assess the diagnostic utility of CE for complex hemoglobinopathies.
Main Methods:
- Retrospective analysis of laboratory results from July 2009 to July 2012.
- Utilized Capillary Electrophoresis (CE) and High-Performance Liquid Chromatography (HPLC) for hemoglobinopathy diagnosis.
- Employed molecular analysis for diagnosing alpha-thalassemia-1 (SEA and Thai types), Hb Constant Spring (CS), and Hb Paksé.
Main Results:
- CE demonstrated superior detection of Hb H-Constant Spring/Paksé-E disease compared to HPLC.
- Hb Bart's and Hb CS/Paksé peaks were consistently observed and quantified by CE in all seven positive samples.
- HPLC could only quantify Hb CS/Paksé levels in 43% of the positive samples, highlighting its limitations.
Conclusions:
- Capillary electrophoresis (CE) is superior to HPLC for the detection and quantification of Hb H-Constant Spring/Paksé-E disease.
- The findings support the use of CE in diagnostic, counseling, and prevention programs for these specific hemoglobinopathies.
- Accurate identification of complex hemoglobin variants through CE improves patient care and genetic counseling.
Abstract:
A capillary electrophoresis (CE) method has been proven to be superior to a high performance liquid chromatography (HPLC) method in the detection of Hb H-Constant Spring/Paksé [Hb H (β4) Hb CS or α142, Term→Gln, TAA>CAA (α2)]/Paksé [α142, Term→Tyr, TAA>TAT (α2)]. It also has the ability to quantify Hb Bart's (γ4). The aim of this study is to analyze the efficacy of the CE and HPLC in the detection of Hb H (β4)-CS/Paksé-E [β26(B8)Glu→Lys, GAG>AAG] disease. The laboratory results from July 2009 to July 2012 were reviewed at the Thalassemia Laboratory of the Associated Medical Sciences Clinical Service Center, Chiang-Mai University, Chiang-Mai, Thailand. The HPLC or CE method was used for the diagnosis of β-thalassemia (β-thal) and hemoglobinopathies, and molecular analysis was used for the diagnosis of α-thalassemia-1 (α-thal-1) Southeast Asian (SEA) and Thai type deletions, Hb CS and Hb Paksé. Hb H-CS-E was found in six samples and Hb H-Paksé-E was found in one sample, respectively. On the capillary electrophoregram, peaks of Hb Bart's and Hb CS/Paksé were observed in all samples with the mean levels at 2.4 and 1.0%, respectively. These peaks were also presented on the HPLC chromatogram. However, the Hb CS/Paksé level could be quantified in only three of these seven (43.0%) samples. Therefore, CE was proven to be superior to HPLC in the detection of Hb H-CS/Paksé-E disease, which will assist in diagnostic, counseling and prevention programs for these diseases.
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