Evaluation of a bead-based enzyme immunoassay for the rapid detection of osteocalcin in human serum.
BACKGROUND
Circulating osteocalcin is a well-known marker for bone formation, but none of the commercial kits currently available can be used in automated systems. Here we present the first semiautomated assay for human serum osteocalcin.


METHODS
Polystyrene beads were coated with antibodies against the COOH terminus of osteocalcin and used in the COBAS((R)) EIA System. Osteocalcin was detected with peroxidase-conjugated antibodies against the osteocalcin NH(2) terminus.


RESULTS
The time required to analyze an unknown sample was 60 min, with a lower detection limit of 4.5 microg/L and a linear dose-response curve between 4.5 and 100 microg/L. The intraassay imprecision (CV) was 5-8% (n = 21); the interassay variation was 6-9% (n = 14). In samples from human volunteers and patients, data generated with the newly developed assay were comparable to those obtained with standard microtiter plate-based assays.


CONCLUSIONS
The coated beads assay may be implemented on fully automated analyzers, which not only may further reduce imprecision but may also substantially increase the applicability of osteocalcin as a marker for bone metabolism in the routine clinical setting.
Crăciun
A M
AM
Department of Biochemistry and Cardiovascular Research Institute, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands.
Vermeer
C
C
Eisenwiener
H G
HG
Drees
N
N
Knapen
M H
MH
D000328:Adult; D000367:Age Factors; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000097:blood; D001842:Bone and Bones / Q000378:metabolism; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods; D008297:Male; D008863:Microspheres; D008875:Middle Aged; D015675:Osteocalcin / Q000097:blood* / Q000276:immunology; D015663:Osteoporosis, Postmenopausal / Q000097:blood / Q000188:drug therapy; D011137:Polystyrenes; D011933:Reagent Kits, Diagnostic; D012737:Sex Factors
D015415:Biomarkers; D011137:Polystyrenes; D011933:Reagent Kits, Diagnostic; D015675:Osteocalcin