Immunonephelometric assay of vitamin D-binding protein.
An automated immunonephelometric assay was developed to measure vitamin D-binding protein (DBP) in serum. The assay is described for the Behring Nephelometer System, which uses rabbit anti-DBP antiserum and purified human DBP. The detection limit is 0.05 g/L (0.86 mumol/L), and the working range is less than or equal to 1.60 g/L (27.59 mumol/L). Intra- and interassay CVs of 2.0% and 2.8-3.8% compare favorably with alternative methods. When results were compared with those from a immunoradiometric assay, the correlation coefficient was 0.976 (P less than 0.001), and the regression equation [y = 0.866 +/- 0.085x + 0.05 (Syx = 0.042, n = 42)] identified a negative bias. Analysis indicated that both methods appeared to contribute equally to the bias. Although the assay was relatively free from analytical interference, falsely increased values were noted in severely lipemic specimens and in frozen specimens. Interference may be minimized by inclusion of Supplementary Precipitation reagent in a modified assay protocol. The range of concentrations expected in clinical samples was established from normal subjects [0.32-0.46 g/L (5.52-7.93 mumol/L), n = 28], pregnant subjects [0.51-0.70 g/L (8.79-12.07 mumol/L), n = 13], and subjects with liver diseases [0.12-0.33 g/L (2.07-5.69 mumol/L), n = 18].
Haughton
M A
MA
Department of Biochemistry, Royal North Shore Hospital, St. Leonards, NSW, Australia.
Mason
R S
RS
D002138:Calibration; D005260:Female; D006801:Humans; D009391:Nephelometry and Turbidimetry / Q000295:instrumentation / Q000379:methods*; D011247:Pregnancy; D011863:Radioimmunoassay; D012016:Reference Values; D015203:Reproducibility of Results; D014809:Vitamin D-Binding Protein / Q000097:blood*
D014809:Vitamin D-Binding Protein