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Automated Capillary Electrophoresis in the Screening for Hemoglobinopathies

Protocol
First Online:
Part of the Methods in Molecular Biology book series (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.

Key words

Hemoglobin Variant Hemoglobinopathy Thalassemia Sickle cell disease Capillary electrophoresis Automation Neonatal screening 
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References

  1. 1.
    Higgs DR, Engel JD, Stamatoyannopoulos G (2012) Thalassaemia. Lancet 379:373–383PubMedCrossRefGoogle Scholar
  2. 2.
    Rees DC, Williams TN, Gladwin MT (2010) Sickle-cell disease. Lancet 376:2018–2031PubMedCrossRefGoogle Scholar
  3. 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 27:3108–3124PubMedCrossRefGoogle Scholar
  4. 4.
    Clarke GM, Higgins TN (2000) Laboratory investigation of hemoglobinopathies and thalassemias: review and update. Clin Chem 46:1284–1290PubMedGoogle Scholar
  5. 5.
    Wajcman H, Prehu C, Bardakdjian-Michau J, Prome D, Riou J, Godart C et al (2001) Abnormal hemoglobins: laboratory methods. Hemoglobin 25:169–181PubMedCrossRefGoogle Scholar
  6. 6.
    Bain BJ (2011) Haemoglobinopathy diagnosis: algorithms, lessons and pitfalls. Blood Rev 25:205–213PubMedCrossRefGoogle Scholar
  7. 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 62:49–52PubMedCrossRefGoogle Scholar
  8. 8.
    Chen FT, Liu CM, Hsieh YZ, Sternberg JC (1991) Capillary electrophoresis - a new clinical tool. Clin Chem 37:14–19PubMedGoogle Scholar
  9. 9.
    Jenkins MA, Hendy J, Smith IL (1997) Evaluation of hemoglobin A2 quantitation assay and hemoglobin variant screening by capillary electrophoresis. J Capillary Electrophor 4:137–143PubMedGoogle Scholar
  10. 10.
    Shihabi ZK, Hinsdale ME, Daugherty HK Jr (2000) Hemoglobin A2 quantification by capillary zone electrophoresis. Electrophoresis 21:749–752PubMedCrossRefGoogle Scholar
  11. 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 45:237–243PubMedGoogle Scholar
  12. 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 23:97–109PubMedCrossRefGoogle Scholar
  13. 13.
    Gulbis B, Fontaine B, Vertongen F, Cotton F (2003) The place of capillary electrophoresis techniques in screening for haemoglobinopathies. Ann Clin Biochem 40:659–662PubMedCrossRefGoogle Scholar
  14. 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 43:2137–2142PubMedGoogle Scholar
  15. 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 719:47–54PubMedCrossRefGoogle Scholar
  16. 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 44:340–345PubMedCrossRefGoogle Scholar
  17. 17.
    Cotton F, Malaviolle X, Vertongen F, Gulbis B (2009) Evaluation of an automated capillary electrophoresis system in the screening for hemoglobinopathies. Clin Lab 55:217–221PubMedGoogle Scholar
  18. 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 152:178–184PubMedCrossRefGoogle Scholar
  19. 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 131:42–48PubMedCrossRefGoogle Scholar
  20. 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 131:357–362PubMedCrossRefGoogle Scholar
  21. 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):362–368PubMedCrossRefGoogle Scholar
  22. 22.
    Higgins T, Mack M, Khajuria A (2009) Comparison of two methods for the quantification and identification of hemoglobin variants. Clin Biochem 42:701–705PubMedCrossRefGoogle Scholar
  23. 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 130:824–831PubMedCrossRefGoogle Scholar
  24. 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 33:424–433PubMedCrossRefGoogle Scholar
  25. 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 47:1423–1432PubMedCrossRefGoogle Scholar
  26. 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 43:1345–1350PubMedCrossRefGoogle Scholar
  27. 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 33:533–539PubMedGoogle Scholar
  28. 28.
    Wolff F, Cotton F, Gulbis B (2012) Screening for haemoglobinopathies on cord blood: a report of laboratory and clinical experience. J Med Screen 19:116–122Google Scholar
  29. 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 42:1780–1785PubMedCrossRefGoogle Scholar
  30. 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 44:406–411PubMedCrossRefGoogle Scholar
  31. 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 90:741–746PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  1. 1.Department of Clinical Chemistry, Erasme HospitalUniversité Libre de BruxellesBrusselsBelgium
  2. 2.Laboratory of Clinical Chemistry, CHU BrugmannBrusselsBelgium
  3. 3.Department of Clinical Chemistry, Hôpital ErasmeUniversité Libre de BruxellesBrusselsBelgium

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