Macro Vitamine B12

How are false elevations of vitamin B12 levels measured?

False increased levels are measure due to presence of macro vitamin B12
Measurement of vitamin B12 performed by analyzers like Roche Cobas or Abott Architect or DiaSorin LIAISON XL in the clinical laboratory may not always accurately reflect real vitamin B12 levels.

Yes, CLIA analyzer Maglumi 800, Snibe uses a pretreatement that eliminates the macro B12 interference.
Due to the presence of Macro B12 CLIA measurement measures false values.
The assays for vitamin B12 were performed in our laboratory using the UniCelR DxI 800 Cbl assay (Beckman Coulter, Brea, CA, USA), and another assay was performed in a different laboratory using the Elecsys E170 Cbl assay (Roche Diagnostics Corp, Indianapolis, IN, USA). Despite high vitamin B12 levels in repeated assays owing to a very strong suspicion of pernicious anemia, further investigation was performed to establish vitamin B12 deficiency and parenteral vitamin B12 replacement was initiated. Cyanocobalamin was administered by intramuscular injection at an initial dose of 1,000 mcg once per day for 1 week and followed by 1,000 mcg once per week. Approximately 2 weeks after the supplementation was initiated, clinical and hematological recovery was observed.

Holotranscobalamin (holo-TC), also known as active B12, is the only form of vitamin B12 that is taken up and used by the cells in the body. It accounts for approximately 10% of the circulating vitamin B12 and is the earliest marker showing vitamin B12 depletion [1,2].
Vitamin B12 deficiency is generally suspected based on related symptoms, clinical findings, and laboratory results and is confirmed by measuring vitamin B12 levels. However, current vitamin B12 measurement methods may miss the lack of vitamin B12 in some cases. These methods, based on competitive binding luminescence assays, have been used since 1990. The assay uses binding to intrinsic factor (IF) following dissociation from the binding proteins, with a readout based on the remaining amount of unbound IF. The main problem with these assays is caused by the presence of IF antibodies in the test sample. IF antibodies may bind the test IF reagent and if there is a failure in the denaturation step intended to denature IF-blocking antibodies, spuriously normal or increased vitamin B12 levels can be measured [3,4]. Low vitamin B12 levels can be measured as false normal or false high, especially in pernicious anemia, due to excessive amounts of anti-intrinsic factor antibodies present in the serum [5,6,7].
In the light of data from the available literature, a normal or high vitamin B12 measurement does not exclude vitamin B12 deficiency in cases when vitamin B12 deficiency is suspected. In such an instance, holo-TC and/or metabolic tests, such as homocysteine or methylmalonic acid, may be considered for patients for whom there is a high suspicion of pernicious anemia in the absence of a low vitamin B12 level. Additionally, an alternate approach involves providing vitamin B12 treatment and confirming or eliminating vitamin B12 deficiency according to the response status.


1. Nexø E, Andersen J. Unsaturated and cobalamin saturated transcobalamin I and II in normal human plasma. Scand J Clin Lab Invest. 1977;37:723–728. [PubMed] [Google Scholar]
2. Nexo E, Hoffmann-Lücke E. Holotranscobalamin, a marker of vitamin B-12 status: analytical aspects and clinical utility. Am J Clin Nutr. 2011;94:359S–365S. [PMC free article] [PubMed] [Google Scholar]
3. Oberley MJ, Yang DT. Laboratory testing for cobalamin deficiency in megaloblastic anemia. Am J Hematol. 2013;88:522–526. [PubMed] [Google Scholar]
4. Vlasveld LT, van’t Wout JW, Meeuwissen P, Castel A. High measured cobalamin (vitamin B12) concentration attributable to an analytical problem in testing serum from a patient with pernicious anemia. Clin Chem. 2006;52:157–158. discussion 158-9. [PubMed] [Google Scholar]
5. van Rossum AP, Vlasveld LT, Castel A. Falsely elevated cobalamin concentration in multiple assays in a patient with pernicious anemia: a case study. Clin Chem Lab Med. 2013;51:e217–e219. [PubMed] [Google Scholar]
6. Shah DR, Daver N, Borthakur G, Hirsch-Ginsberg C, Oo TH. Pernicious anemia with spuriously normal vitamin B12 level might be misdiagnosed as myelodysplastic syndrome. Clin Lymphoma Myeloma Leuk. 2014;14:e141–e143. [PubMed] [Google Scholar]
7. Shishido T, Hiroshima Y, Uematsu N, et al. Successful treatment with mecobalamin in a pernicious anemia patient presenting with false-normal serum vitamin B12. Rinsho Ketsueki. 2018;59:675–681. [PubMed] [Google Scholar]

Djoumana Ounas

Djoumana Ounas

Pharmacist Assistant Professor in Analytical Chemisty. Consultance Expertise and Reglatory Biotech Support.

want a datasheet ?

want A Quote ?