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Screening for co-existence of α-thalassemia in β-thalassemia and in HbE heterozygotes via an enzyme-linked immunosorbent assay for Hb Bart’s and embryonic ζ-globin chain

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Abstract

We sought to demonstrate the ability of levels of Hb Bart’s and ζ-globin chain quantified by enzyme-linked immunosorbent assay (ELISA) in detecting α-thalassemia in β-thalassemia and HbE heterozygotes. We developed an in-house sandwich ELISA method using monoclonal antibodies (mAbs) to Hb Bart’s and ζ-globin chain, and quantified levels of Hb Bart’s and ζ-globin chain in 172 and 223 anonymous blood samples of β-thalassemia and HbE heterozygotes, respectively. Genotypes of α-thalassemia 1, β-thalassemia were identified, and HbE allele was confirmed using a newly developed multiplex allele-specific PCR. The in-house sandwich ELISA method detected Hb Bart’s in 6.4% of β-thalassemia heterozygotes, of which 5.2% showed detectable amounts of the ζ-globin chain. 15.2% of individuals heterozygous for HbE showed a detectable amount of Hb Bart’s, and the ζ-globin chain was detected in 11.2% of this cohort. All samples having detectable amounts of Hb Bart’s and the ζ-globin chain were verified to be SEA-type α-thalassemia 1. ELISA-quantified Hb Bart’s and ζ-globin chain levels can be used to detect double heterozygosity of α- and β-thalassemia and of α-thalassemia and HbE. This strategy may be useful in screening for co-existence of α-thalassemia in β-thalassemia and in HbE heterozygotes, particularly in countries where α-, β-thalassemia and HbE are endemic.

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Acknowledgments

This study was supported by the NSTDA Research Chair Grant, National Sciences and Technology Development Agency (Thailand) and the National Research University Project under Thailands Office of the Higher Education Commission. We thank Dr. Supansa Pata and Ms. Jirawan Poorahong for the technical assistant. We also thank Dr. Dale E. Taneyhill for proofreading the manuscript.

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Authors and Affiliations

  1. Division of Clinical Microscopy, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand

    Thanusak Tatu & Surasit Suwannasin

  2. Laboratory Hematology Unit, Department of Clinical Pathology, Lampang Central Hospital, Amphor Muang, Lampang, 52000, Thailand

    Tiemjan Kiewkarnkha

  3. Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand

    Surakit Khuntarak, Sakdinan Khamrin & Watchara Kasinrerk

  4. Faculty of Associated Medical Sciences, Biomedical Technology Research Center, National Center for Genetic Engineering and Biotechnology, National Sciences and Technology Development Agency, Chiang Mai University, Chiang Mai, 50200, Thailand

    Watchara Kasinrerk

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  1. Thanusak Tatu
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  2. Tiemjan Kiewkarnkha
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  3. Surakit Khuntarak
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  5. Surasit Suwannasin
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  6. Watchara Kasinrerk
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Correspondence to Thanusak Tatu or Watchara Kasinrerk.

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Tatu, T., Kiewkarnkha, T., Khuntarak, S. et al. Screening for co-existence of α-thalassemia in β-thalassemia and in HbE heterozygotes via an enzyme-linked immunosorbent assay for Hb Bart’s and embryonic ζ-globin chain. Int J Hematol 95, 386–393 (2012). https://doi.org/10.1007/s12185-012-1039-4

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  • DOI: https://doi.org/10.1007/s12185-012-1039-4

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