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Macro vitamin B12: an underestimated threat

  • Reza Soleimani ORCID logo , Julien Favresse , Tatiana Roy , Damien Gruson EMAIL logo und Catherine Fillée
Veröffentlicht/Copyright: 30. Oktober 2019
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Abstract

Background

The correct identification of the macro-B12 interference (macroforms) is paramount to avoid potential erroneous clinical decisions. Our objectives were to determine whether immunoassays are affected by the presence of macro-B12 and to validate a polyethylene glycol (PEG) precipitation procedure to detect it.

Methods

Sixty-two serum samples obtained from healthy volunteers were analyzed to determine recovery and reference intervals (RIs) following PEG precipitation. Thereafter, 50 serum samples with very high levels of B12 (>1476 pmol/L) were randomly selected to search for macro-B12 interferences. Serum samples obtained from healthy volunteers and related PEG aliquots were analyzed on a Cobas® immunoassay. Patients’ samples were analyzed on both Cobas® and Architect® immunoassays. Finally, samples suspected to contain macro-B12 were analyzed by size-exclusion chromatography (SEC) to confirm the presence of macro-B12.

Results

Recovery and post-PEG RIs determined on a Cobas 8000® in healthy volunteers ranged from 68.3% to 108.4% and from 122.1 to 514.4 pmol/L, respectively. Fifteen samples (30%) were found to show macro-B12 while using the recovery criteria, and nine samples (18%) while using the post-PEG RI. The other immunoassay ran on the Architect i2000® was also affected by the presence of macro-B12. Size-exclusion chromatography studies confirmed the presence of macro-B12 (immunoglobulin-B12 complexes).

Conclusions

The prevalence of macro-B12 in elevated B12 samples is high. We suggest to systematically screen for the presence of macro-B12 with PEG precipitation procedure in samples with elevated B12 levels to avoid potential misdiagnosis or harmful clinical consequences.


Corresponding author: Prof. Damien Gruson, Clinical Chemistry Service, Cliniques Universitaires Saint Luc, Rosalind Franklin Building, 49 Avenue Mounier, 1200 Brussels, Belgium; Department of Laboratory Medicine, Cliniques Universitaires St-Luc and Université catholique de Louvain, Brussels, Belgium; and Research Pole of Endocrinology, Diabetes and Nutrition, Institute of Experimental and Clinical Research, Cliniques Universitaires St-Luc and Université catholique de Louvain, Brussels, Belgium, Phone: +32-(0)2-7646747, Fax: +32-(0)2-7646932
aReza Soleimani and Julien Favresse contributed equally to this work.

Acknowledgments

The authors would like to thank the laboratory technicians, Mrs. Corinne Martin, Mr. Jürgen Lohmann, Mr. Mehdi Mancini, Mr. Matteo Murer, Mrs. Hajar Latrache, Mr. Helder Dos Santos and Mr. Jacques Vanham as well as the laboratories residents, Mrs. Anke Stoefs, Mr. Samy Mzougui and Antoine Mairesse, without whom this work would not have been possible. The authors also acknowledge Roche Diagnostics for performing the size-exclusion chromatography.

  1. Author contributions: All the authors have approved the entire content of the submitted manuscript (and any subsequent revised version) and have accepted responsibility for the entire work.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Received: 2019-09-25
Accepted: 2019-10-06
Published Online: 2019-10-30
Published in Print: 2020-02-25

©2020 Walter de Gruyter GmbH, Berlin/Boston

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