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Reliability of hemoglobin A2 value as measured by the Premier Resolution system for screening of β-thalassemia carriers

  • Surada Satthakarn , Kesorn Panyasai , Amphai Phasit and Sitthichai Panyasai EMAIL logo
Published/Copyright: October 17, 2023
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 62 Issue 3

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.

Keywords: capillary electrophoresis; hemoglobinopathies; high-performance liquid chromatography; hemoglobin A2 ; β-thalassemia
Corresponding Author: Dr. Sitthichai Panyasai, Department of Medical Technology, School of Allied Health Sciences, University of Phayao, 19, Phahonyothin Road, Muang Phayao, Phayao, Thailand, E-mail:

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.

  1. 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.

  2. Informed consent: Informed consent was obtained from all individuals included in this study.

  3. 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.

  4. Competing interests: Authors state no conflict of interest.

  5. 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].

  6. 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).

Received: 2023-09-10
Accepted: 2023-09-30
Published Online: 2023-10-17
Published in Print: 2024-02-26

© 2023 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2023-1006
Keywords for this article
capillary electrophoresis; hemoglobinopathies; high-performance liquid chromatography; hemoglobin A2; β-thalassemia

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Unraveling the mystery of blood groups and COVID-19
  4. Reviews
  5. Serum hepcidin levels in chronic liver disease: a systematic review and meta-analysis
  6. Platelet distribution width (PDW) as a significant correlate of COVID-19 infection severity and mortality
  7. Mini Reviews
  8. ABO blood group-related mechanism of infection of SARS-CoV-2: an overview of systematic reviews
  9. Opinion Paper
  10. Personalized laboratory medicine in the digital health era: recent developments and future challenges
  11. Guidelines and Recommendations
  12. Algorithm of differential diagnosis of anemia involving laboratory medicine specialists to advance diagnostic excellence
  13. General Clinical Chemistry and Laboratory Medicine
  14. Rescaling creatinine makes GFR estimation equations generally applicable across populations – validation results for the Lund-Malmö equation in a French cohort of sub-Saharan ancestry
  15. Periodic verification of results’ comparability between several analyzers: experience in the application of the EP31-A-IR guideline
  16. Machine learning to optimize cerebrospinal fluid dilution for analysis of MRZH reaction
  17. Diagnostic performance of automated red cell parameters in predicting bone marrow iron stores
  18. Reliability of hemoglobin A2 value as measured by the Premier Resolution system for screening of β-thalassemia carriers
  19. Amino acid sequence homology of monoclonal serum free light chain dimers and tissue deposited light chains in AL amyloidosis: a pilot study
  20. Development of high-performance point-of-care aqueous VEGF detection system and proof-of-concept validation in RVO patients
  21. Detection rate of IGF-1 variants and their implication to protein binding: study of over 240,000 patients
  22. Analysis of a second-tier test panel in dried blood spot samples using liquid chromatography-tandem mass spectrometry in Catalonia’s newborn screening programme
  23. Targeted quantitative lipidomic uncovers lipid biomarkers for predicting the presence of compensated cirrhosis and discriminating decompensated cirrhosis from compensated cirrhosis
  24. Reference Values and Biological Variations
  25. Establishment of reference intervals for free light chains and immunoglobulins in Saudi population
  26. Cancer Diagnostics
  27. A predictive and prognostic model for surgical outcome and prognosis in ovarian cancer computed by clinico-pathological and serological parameters (CA125, HE4, mesothelin)
  28. M-protein diagnostics in multiple myeloma patients using ultra-sensitive targeted mass spectrometry and an off-the-shelf calibrator
  29. Cardiovascular Diseases
  30. Bioactive adrenomedullin (bio-ADM) is associated with endothelial dysfunction in infants and children with complex congenital heart disease undergoing open-heart surgery on cardiopulmonary bypass
  31. Infectious Diseases
  32. Monocyte distribution width as an early predictor of short-term outcome in adult patients with sepsis
  33. Analytical and clinical evaluations of SNIBE Maglumi chemiluminescent immunoassay for the detection of SARS-CoV-2 antigen in salivary samples
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  36. Managing the Quality Control of multiple instruments
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  38. Assessment of the lipemia index determined by the Atellica CH 930 analyzer for the detection of monoclonal immunoglobulins
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  41. Early detection of peripheral invasive candidiasis further to cytographic interferences in Sysmex XN-9000 hematology analyzer
  42. Congress Abstracts
  43. Annual meeting of the Royal Belgian Society of Laboratory Medicine: “Symphony of the Heart”
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