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INTRODUCTION
surement of holoTC is also likely to be valuable for populationwide screening of vitamin B-12 status.
HoloTC AND OTHER MARKERS OF VITAMIN B-12
STATUS
From the Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus, Denmark.
2
Presented at the conference NHANES Monitoring of Biomarkers of
Folate and Vitamin B-12 Status: a Roundtable Review, held in Rockville,
MD, 1516 July 2010.
3
The present review is, to a large extent, based on results from a European
Union demonstration project (HoloTC: Early B12 Marker, QLK3-CT-200201775, 20022006) with participants from Denmark: Ebba Nexo (coordinator), Tore F Hardlei, Anne-Mette Hvas, Anne Louise Morkbak], England
(Robert Clarke), Ireland (Anne Molloy, John M Scott), Norway (Jorn
Schneede, Per Magne Ueland), and Scotland (Edward Valente).
4
Supported by the Danish Medical Research Council and the Lundbeck
Foundation. The National Center for Health Statistics of the Centers for
Disease Control and Prevention and the Office of Dietary Supplements of
the National Institutes of Health cosponsored and supported the roundtable.
5
Address correspondence and reprint requests to E Nexo, Department of
Clinical Biochemistry, Aarhus University Hospital, Aarhus, Norrebrogade
44, DK 8000 Aarhus C, Denmark. E-mail: e.nexo@dadlnet.dk.
doi: 10.3945/ajcn.111.013458.
Am J Clin Nutr doi: 10.3945/ajcn.111.013458. Printed in USA. 2011 American Society for Nutrition
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NEXO AND HOFFMANN-LU
Three different types of holoTC measurement methods are available and, as described below, we have shown that these methods
give reasonably similar values. The first approach to be published
combined ionic precipitation of transcobalamin with measurement of the amount of vitamin B-12 trapped in the precipitate
(1820).Lindemans etal(21) improved thisapproach withthe use of
antibodies directed to transcobalamin rather than ionic separation,
and Ulleland et al (22) developed a version of this approach that
involved measurement of trapped vitamin B-12 by an isotope
dilution assay, the holoTC-radioimmunoassay (RIA). In another
version, Refsum et al (9) measured vitamin B-12 with the use of
a microbiological method. Yet another approach is to remove apotranscobalamin with vitamin B-12coated beads followed by
enzyme-linked immunosorbent assay measurement of the holoTC
that remains in the supernatant fluid (23).
A multicenter study that involved 4 European laboratories
evaluated the holoTC-RIA (calibration curve: 10160 pmol/L) and
the holoTCenzyme-linked immunosorbent assay (calibration curve:
1.6100 pmol/L) (24). The results from the study showed similar
values across laboratories and across the methods tested. The longterm imprecision was 6% for the enzyme-linked immunosorbent
assay and .10% for the RIA (24). The RIA assay results were
also similar to the results that Refsum et al (9) obtained with the use
of the microbiological assay (calibration curve: 1.3624.5 pmol/L;
imprecision: 47%).
Today, the holoTC-RIA has been replaced by an assay that uses
holoTC-specific monoclonal antibodies (25). On the AxSYM
platform (Abbott Labs, North Chicago, IL), the method has
a measurement range of 3100 pmol/L and a total imprecision of
69%. The results obtained by this method compare well with
those of the holoTC-RIA (26, 27).
HoloTC DETERMINANTS
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Transcobalamin genotypes
Transcobalamin occurs in many genotypes (39), and several
authors have clearly documented that the genotype present,
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NEXO AND HOFFMANN-LU
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TABLE 1
Selected reference intervals for holotranscobalamin (holoTC) with the use
of the most common assays for holoTC measurement1
Method2
Sample
lL
400
Radioimmunoassay
Microbiology
ELISA
Direct
,150
100
200
Reference
range
Reference
pmol/L
24160
37170
42160
40150
19130
36220
105
303
500
137
292
276
22
28
9
23
26
50
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Nationality
Age
Limits for
MMA (tHcy)
AUC for
holoTC
AUC for
vitamin B-12
Reference
Danish
English
English
German
German
German
German, Dutch
y
.18
.65
.65
1898
1898
892
2173
lmol/L
.0.75 (.15)
.0.75
.0.45
.0.40
.0.40
.0.30
.0.27
0.90
0.87
0.80
0.66
0.71
0.88
0.85
0.79
0.73
0.72
0.60
0.84
32
56
56
59
59
58
57
1
The limits for MMA and tHcy indicate the cutoffs that were used to classify the patients as vitamin B-12 deficient. The researchers assessed the results
with the use of receiver operating curves. An area under the curve (AUC) of 1.0 indicates a perfect test, and an AUC of 0.5 indicates a useless test. MMA,
methylmalonic acid; tHcy, homocysteine.
the tests will be able to classify individuals to have a low, but not
absent, capacity for absorption of the vitamin.
CONCLUSIONS
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