Skip to main content

Automated Capillary Electrophoresis in the Screening for Hemoglobinopathies

  • Protocol
  • First Online:
Capillary Electrophoresis of Biomolecules

Part of the book series: Methods in Molecular Biology ((MIMB,volume 984))

Abstract

Hemoglobinopathies are genetic disorders of globin chains characterized by the decreased expression of α- or β-globin chains (thalassemias) or by the synthesis of an abnormal protein (hemoglobin variants in, e.g., sickle cell disease). The screening of most hemoglobinopathies relies, together with hematological results and clinical elements, on the separation and quantification of normal and abnormal hemoglobin fractions. Gel electrophoresis, isoelectric focusing, and HPLC have been the methods of choice for many years. For about 20 years, capillary electrophoresis has appeared as a strong alternative method. Since the early 2000s, automated instruments are commercially available for the analysis of Hb fractions in adult patients but also for neonatal screening.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Higgs DR, Engel JD, Stamatoyannopoulos G (2012) Thalassaemia. Lancet 379:373–383

    Article  PubMed  CAS  Google Scholar 

  2. Rees DC, Williams TN, Gladwin MT (2010) Sickle-cell disease. Lancet 376:2018–2031

    Article  PubMed  CAS  Google Scholar 

  3. Wang J, Zhou S, Huang W, Liu Y, Cheng C, Lu X, Cheng J (2006) CE-based analysis of hemoglobin and its applications in clinical analysis. Electrophoresis 27:3108–3124

    Article  PubMed  CAS  Google Scholar 

  4. Clarke GM, Higgins TN (2000) Laboratory investigation of hemoglobinopathies and thalassemias: review and update. Clin Chem 46:1284–1290

    PubMed  CAS  Google Scholar 

  5. Wajcman H, Prehu C, Bardakdjian-Michau J, Prome D, Riou J, Godart C et al (2001) Abnormal hemoglobins: laboratory methods. Hemoglobin 25:169–181

    Article  PubMed  CAS  Google Scholar 

  6. Bain BJ (2011) Haemoglobinopathy diagnosis: algorithms, lessons and pitfalls. Blood Rev 25:205–213

    Article  PubMed  Google Scholar 

  7. Gulbis B, Cotton F, Ferster A, Ketelslegers O, Dresse MF, Rongé-Collard E, Minon JM, Lé PQ, Vertongen F (2009) Neonatal haemoglobinopathy screening in Belgium. J Clin Pathol 62:49–52

    Article  PubMed  CAS  Google Scholar 

  8. Chen FT, Liu CM, Hsieh YZ, Sternberg JC (1991) Capillary electrophoresis - a new clinical tool. Clin Chem 37:14–19

    PubMed  CAS  Google Scholar 

  9. Jenkins MA, Hendy J, Smith IL (1997) Evaluation of hemoglobin A2 quantitation assay and hemoglobin variant screening by capillary electrophoresis. J Capillary Electrophor 4:137–143

    PubMed  CAS  Google Scholar 

  10. Shihabi ZK, Hinsdale ME, Daugherty HK Jr (2000) Hemoglobin A2 quantification by capillary zone electrophoresis. Electrophoresis 21:749–752

    Article  PubMed  CAS  Google Scholar 

  11. Cotton F, Lin C, Fontaine B, Gulbis B, Janssens J, Vertongen F (1999) Evaluation of a capillary electrophoresis method for routine determination of hemoglobins A2 and F. Clin Chem 45:237–243

    PubMed  CAS  Google Scholar 

  12. Lin C, Cotton F, Fontaine B, Gulbis B, Janssens J, Vertongen F (1999) Capillary zone electrophoresis: an additional technique for the identification of hemoglobin variants. Hemoglobin 23:97–109

    Article  PubMed  CAS  Google Scholar 

  13. Gulbis B, Fontaine B, Vertongen F, Cotton F (2003) The place of capillary electrophoresis techniques in screening for haemoglobinopathies. Ann Clin Biochem 40:659–662

    Article  PubMed  CAS  Google Scholar 

  14. Mario N, Baudin B, Aussel C, Giboudeau J (1997) Capillary isoelectric focusing and high-performance cation-exchange chromatography compared for qualitative and quantitative analysis of hemoglobin variants. Clin Chem 43:2137–2142

    PubMed  CAS  Google Scholar 

  15. Lin C, Gulbis B, Delobbe E, Cotton F, Vertongen F (1998) Separation of human globin chains by micellar electrokinetic capillary chromatography. J Chromatogr B Biomed Sci Appl 719:47–54

    Article  PubMed  CAS  Google Scholar 

  16. Louahabi A, Philippe M, Lali S, Wallemacq P, Maisin D (2006) Evaluation of a new Sebia kit for analysis of hemoglobin fractions and variants on the Capillarys system. Clin Chem Lab Med 44:340–345

    Article  PubMed  CAS  Google Scholar 

  17. Cotton F, Malaviolle X, Vertongen F, Gulbis B (2009) Evaluation of an automated capillary electrophoresis system in the screening for hemoglobinopathies. Clin Lab 55:217–221

    PubMed  CAS  Google Scholar 

  18. Winichagoon P, Svasti S, Munkongdee T, Chaiya W, Boonmongkol P, Chantrakul N, Fucharoen S (2008) Rapid diagnosis of thalassemias and other hemoglobinopathies by capillary electrophoresis system. Transl Res 152:178–184

    Article  PubMed  CAS  Google Scholar 

  19. Yang Z, Chaffin CH, Easley PL, Thigpen B, Reddy VV (2009) Prevalence of elevated hemoglobin A2 measured by the CAPILLARYS system. Am J Clin Pathol 131:42–48

    Article  PubMed  CAS  Google Scholar 

  20. Higgins TN, Khajuria A, Mack M (2009) Quantification of HbA2 in patients with and without beta-thalassemia and in the presence of HbS, HbC, HbE, and HbD Punjab hemoglobin variants: comparison of two systems. Am J Clin Pathol 131:357–362

    Article  PubMed  CAS  Google Scholar 

  21. Paleari R, Gulbis B, Cotton F, Mosca A (2012) Interlaboratory comparison of current high-performance methods for HbA2. Int J Lab Hematol 34(4):362–368

    Article  PubMed  CAS  Google Scholar 

  22. Higgins T, Mack M, Khajuria A (2009) Comparison of two methods for the quantification and identification of hemoglobin variants. Clin Biochem 42:701–705

    Article  PubMed  CAS  Google Scholar 

  23. Keren DF, Hedstrom D, Gulbranson R, Ou CN, Bak R (2008) Comparison of Sebia Capillarys capillary electrophoresis with the Primus high-pressure liquid chromatography in the evaluation of hemoglobinopathies. Am J Clin Pathol 130:824–831

    Article  PubMed  CAS  Google Scholar 

  24. Fucharoen G, Srivorakun H, Singsanan S, Fucharoen S (2011) Presumptive diagnosis of common haemoglobinopathies in Southeast Asia using a capillary electrophoresis system. Int J Lab Hematol 33:424–433

    Article  PubMed  CAS  Google Scholar 

  25. Renom G, Mereau C, Maboudou P, Périni JM (2009) Potential of the Sebia Capillarys neonat fast automated system for neonatal screening of sickle cell disease. Clin Chem Lab Med 47:1423–1432

    Article  PubMed  CAS  Google Scholar 

  26. Mantikou E, Harteveld CL, Giordano PC (2010) Newborn screening for hemoglobinopathies using capillary electrophoresis technology: Testing the Capillarys Neonat Fast Hb device. Clin Biochem 43:1345–1350

    Article  PubMed  CAS  Google Scholar 

  27. Murray C, Hall SK, Griffiths P (2011) An evaluation of the Sebia capillarys Neonat Haemoglobin FAST™ system for routine ­newborn screening for sickle cell disease. Int J Lab Hematol 33:533–539

    PubMed  CAS  Google Scholar 

  28. Wolff F, Cotton F, Gulbis B (2012) Screening for haemoglobinopathies on cord blood: a report of laboratory and clinical experience. J Med Screen 19:116–122

    Google Scholar 

  29. Mantikou E, Arkesteijn SG, van Beckhoven JM, Kerkhoffs JL, Harteveld CL, Giordano PC (2009) A brief review on newborn screening methods for hemoglobinopathies and preliminary results selecting beta thalassemia carriers at birth by quantitative estimation of the HbA fraction. Clin Biochem 42:1780–1785

    Article  PubMed  CAS  Google Scholar 

  30. Srivorakun H, Fucharoen G, Changtrakul Y, Komwilaisak P, Fucharoen S (2011) Thalassemia and hemoglobinopathies in Southeast Asian newborns: diagnostic assessment using capillary electrophoresis system. Clin Biochem 44:406–411

    Article  PubMed  CAS  Google Scholar 

  31. Munkongdee T, Pichanun D, Butthep P, Klamchuen S, Chalermpolprapa V, Winichagoon P, Svasti S, Fucharoen S (2011) Quantitative analysis of Hb Bart’s in cord blood by capillary electrophoresis system. Ann Hematol 90:741–746

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Frédéric Cotton .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2013 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Cotton, F., Wolff, F., Gulbis, B. (2013). Automated Capillary Electrophoresis in the Screening for Hemoglobinopathies. In: Volpi, N., Maccari, F. (eds) Capillary Electrophoresis of Biomolecules. Methods in Molecular Biology, vol 984. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-296-4_16

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-296-4_16

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-295-7

  • Online ISBN: 978-1-62703-296-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics