Abstract
Objectives
Accurate quantification of hemoglobin (Hb) A2 is vital for diagnosing β-thalassemia carriers. This study aimed to assess the precision and diagnostic utility of HbA2 measurements using the new high-performance liquid chromatography (HPLC) method, Premier Resolution, in comparison to capillary electrophoresis (CE).
Methods
We analyzed 418 samples, previously identified as A2A by CE, using Premier Resolution-HPLC. We compared the results, established correlations, and determined an optimal HbA2 cutoff value for β-thalassemia screening. Additionally, we prospectively evaluated the chosen cutoff value in 632 samples. Mutations in the β- and α-globin genes were identified using polymerase chain reaction (PCR) techniques and DNA sequencing.
Results
HbA2 levels were consistently higher with Premier Resolution, yet there was a significant correlation with CE in all samples (bias, −0.33; r, 0.991), β-thalassemia (bias, −0.27; r, 0.927), and non-β-thalassemia carriers (bias, −0.36; r, 0.928). An HbA2 cutoff value of ≥4.0 % for β-thalassemia screening achieved 100 % sensitivity and 99.6 % specificity. Further validation yielded sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of 97.3 , 99.8, 97.3, 99.8, and 99.7 %, respectively. We also identified a rare β-Hb variant, Hb La Desirade [HBB:c.389C>T], associated with β-thalassemia and co-inherited with a single α-globin gene.
Conclusions
The Premier Resolution HPLC is a reliable and accurate method for routine β-thalassemia carrier screening, aligning with existing CE methods.
Funding source: Thailand Science Research and Innovation Fund and the University of Phayao, Thailand
Award Identifier / Grant number: FF65-RIM136
Acknowledgments
We would like to thank technicians at the Thalassemia Laboratory in Lampang hospitals for their help and assistance. We also would like to thank Biozen Co., Ltd. for supplying reagents and automation analyzers.
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Research ethics: The research protocal was conducted in accordance with the Declaration of Helsinki (as revised in 2013) and has been approved by the Institutional Review Board (IRB) of the University of Phayao, (1.3/002/65) Phayao, Thailand and Lampang hospital, Lampang, Thailand (EC056/65). Informed consent obtained from all participants.
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Informed consent: Informed consent was obtained from all individuals included in this study.
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Author contributions: Sitthichai Panyasai: Conceptualization, Methodology, Funding acquisition, Writing – original draft, review and editing. Surada Satthakarn: Investigation, Formal analysis, Data Curation, Writing – Original draft preparation. Kesorn Panyasai: Resources, Investigation. Amphai Phasit: Resources, Investigation. All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
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Competing interests: Authors state no conflict of interest.
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Research funding: This study was funded by research grants from the Thailand Science Research and Innovation fund and the University of Phayao, Thailand [grant number FF65-RIM136].
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Data availability: The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
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Supplementary Material
This article contains supplementary material (https://doi.org/10.1515/cclm-2023-1006).
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