Jump to ContentJump to Main Navigation
Show Summary Details
More options …

Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter


IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

SCImago Journal Rank (SJR) 2018: 1.191
Source Normalized Impact per Paper (SNIP) 2018: 1.205

Online
ISSN
1437-4331
See all formats and pricing
More options …
Volume 53, Issue 9

Issues

Two novel haemoglobin variants that affect haemoglobin A1c measurement by ion-exchange chromatography

Michael Bots
  • Department of Clinical Chemistry, Academic Medical Center, University of Amsterdam, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ An K. Stroobants
  • Department of Clinical Chemistry, Academic Medical Center, University of Amsterdam, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Barend Delzenne
  • Department of Clinical Chemistry, Academic Medical Center, University of Amsterdam, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Maarten R. Soeters
  • Department of Endocrinology and Metabolism, Academic Medical Center, University of Amsterdam, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Johan E. de Vries / Cas W. Weykamp
  • MCA Laboratory, Queen Beatrix Hospital, Winterswijk, The Netherlands
  • European Reference Laboratory for Glycohemoglobin, Winterswijk, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Roelf J.C. Norg / Martijn Veldthuis
  • Laboratory of Red Blood Cell Diagnostics, Sanquin Blood Supply Organisation, Amsterdam, The Netherlands
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Rob van Zwieten
  • Corresponding author
  • Laboratory of Red Blood Cell Diagnostics, Sanquin Blood Supply Organisation, Amsterdam, The Netherlands
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2015-06-02 | DOI: https://doi.org/10.1515/cclm-2015-0054

Abstract

Background: Haemoglobin (Hb) variants are well-known factors interfering with accurate HbA1c testing. This report describes two novel Hb variants leading to inappropriate quantification of HbA1c by ion-exchange chromatography.

Methods: Glycated forms of novel Hb variants were recognised in the blood of two patients with diabetes mellitus screened by HbA1c ion-exchange chromatography. Dedicated high-resolution cation-exchange chromatography and subsequent DNA sequencing revealed the exact nature of the variants. Other common techniques for quantifying HbA1c were applied on both samples and haematological parameters were determined to judge possible pathology associated with the novel Hb variants.

Results: A fraction of 15% of abnormal Hb was observed in a 37-year-old female. DNA sequencing revealed a heterozygous mutation in the α1-globin gene, resulting in a leucine-to-phenylalanine amino-acid substitution (HBA1: c.301C>T, p.Leu101Phe). We named this variant Hb Weesp. The other novel variant, Hb Haelen, presented as a 40% fraction in a 63-year-old male and resulted from a heterozygous amino acid substitution in the β-globin gene (HBB: c.335T>C, p.Val112Gly). The presence of both Hb variants resulted in aberrant separation of the Hb components, leading to an inadequate quantification of HbA1c.

Conclusions: Close examination of HbA1c chromatograms revealed two novel, clinically silent Hb variants that interfere with HbA1c quantification. Healthcare providers need to be aware of the potential of such Hb variants when interpreting HbA1c results.

Keywords: diabetes mellitus; glycated haemoglobin; HbA1c; Hb variant; ion-exchange chromatography

References

  • 1.

    Burtis CA, Ashwood ER, Bruns DE, editors. Tietz textbook of clinical chemistry and molecular diagnostics, 7th ed. st. Louis: WB Saunders/Elsevier, 2015.Google Scholar

  • 2.

    Nathan DM, Kuenen J, Borg R, Zheng H, Schoenfeld D, Heine RJ, et al. Translating the A1C assay into estimated average glucose values. Diabetes Care 2008;31:1473–8.CrossrefWeb of ScienceGoogle Scholar

  • 3.

    Bunn HF. Practicing biochemistry without a license. J Biol Chem 2013;288:5062–71.Web of ScienceGoogle Scholar

  • 4.

    Gallagher EJ, Le Roith D, Bloomgarden Z. Review of hemoglobin A1c in the management of diabetes. J Diabetes 2009;1:9–17.Google Scholar

  • 5.

    Little RR, Sacks DB. HbA1c: how do we measure it and what does it mean? Curr Opin Endocrinol Diabetes Obes 2009;16:113–8.CrossrefGoogle Scholar

  • 6.

    Little RR, Roberts WL. A review of variant hemoglobins interfering with hemoglobin A1c measurement. J Diabetes Sci Technol 2009;3:446–51.CrossrefGoogle Scholar

  • 7.

    Rhea JM, Molinaro R. Pathology consultation on HbA1c methods and interferences. Am J Clin Pathol 2013;141:5–16.Web of ScienceGoogle Scholar

  • 8.

    Bry L, Chen PC, Sacks DB. Effects of hemoglobin variants and chemically modified derivatives on assays for glycohemoglobin. Clin Chem 2001;47:153–63.Google Scholar

  • 9.

    Sacks DB. Hemoglobin A1c in diabetes: panacea or pointless? Diabetes 2012;62:41–3.Web of ScienceGoogle Scholar

  • 10.

    Van Zwieten R, Veldthuis M, Delzenne B, Berghuis J, Groen J, Ait Ichou F, et al. Hemoglobin analyses in the Netherlands reveal more than 80 different variants including six novel ones. Hemoglobin 2014;38:1–7.CrossrefPubMedWeb of ScienceGoogle Scholar

  • 11.

    Tan AS, Quah TC, Low PS, Chong SS. A rapid and reliable 7-deletion multiplex polymerase chain reaction assay for alpha-thalassemia. Blood 2001;98:250–1.Google Scholar

  • 12.

    Bain BJ, Bates I, Laffan ME, Lewis SM, editors. Dacie and Lewis. Practical haematology, 11th ed. London: Churchill Livingsone/Elsevier, 2012.Google Scholar

  • 13.

    Cohen RM, Franco RS, Khera PK, Smith EP, Lindsell CJ, Ciraolo PJ, et al. Red cell life span heterogeneity in hematologically normal people is sufficient to alter HbA1c. Blood 2008;112:4284–91.Web of ScienceGoogle Scholar

  • 14.

    Tarim Ö, Küçükerdogan A, Günay Ü, Eralp Ö, Ercan I. Effects of iron deficiency anemia on hemoglobin A1c in type 1 diabetes mellitus. Pediatr Int 1999;41:357–62.Google Scholar

  • 15.

    Little RR, Rohlfing CL. The long and winding road to optimal HbA1c measurement. Clin Chim Acta 2013;418:63–71.Web of ScienceGoogle Scholar

  • 16.

    Miyazaki A, Nakanishi T, Shimizu A, Hisamitsu H. First case of a single heterozygote of an abnormal hemoglobin, Hb Stanmore, [beta111(G13)Val–>Ala]. J. Chromatogr B Analyt Technol Biomed Life Sci 2003;792:23–31.Google Scholar

  • 17.

    Carbone V, Salzano AM, Pagnini D, Papa G, Libertino R, Pagano L, et al. The mutation associated with Hb Peterborough [beta111(G13)Val–>Phe] originated from Southern Italy. Hemoglobin 2000;24:227–37.Google Scholar

About the article

Corresponding author: Rob van Zwieten, Laboratory of Red Blood Cell Diagnostics, Sanquin Blood Supply Organization, Plesmanlaan 125, 1066 CX Amsterdam, The Netherlands, Phone: +31 20 512 3317, Fax: +31 20 512 3310, E-mail:


Received: 2015-01-16

Accepted: 2015-04-29

Published Online: 2015-06-02

Published in Print: 2015-08-01


Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), Volume 53, Issue 9, Pages 1465–1471, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2015-0054.

Export Citation

©2015 by De Gruyter.Get Permission

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

[1]
Tomomi Hatayama, Fumio Umeda, Teruaki Yamauchi, and Hiroshi Ideguchi
Diabetology International, 2019
[2]
Clayton R. Wilburn, David W. Bernard, Arthur W. Zieske, Julia Andrieni, Tara Miller, and Ping Wang
American Journal of Clinical Pathology, 2017, Volume 147, Number 6, Page 589
[3]
Haval Surchi and Rustam Rea
Practical Diabetes, 2017, Volume 34, Number 1, Page 28
[4]
Ayşegül Uğur Kurtoğlu, Esin Eren, Özgür Erkal, Erdal Kurtoğlu, and Necat Yilmaz
Clinica Chimica Acta, 2017, Volume 465, Page 80
[6]
Weiping Jia
Expert Review of Molecular Diagnostics, 2016, Volume 16, Number 3, Page 343
[7]
Philippe Gillery
Clinical Chemistry and Laboratory Medicine (CCLM), 2015, Volume 53, Number 9

Comments (0)

Please log in or register to comment.
Log in