Development of an automated turbidimetric immunoassay for quantification of bovine serum immunoglobulin G.
OBJECTIVE
To develop an automated turbidimetric immunoassay (TIA) for measurement of bovine IgG.


METHODS
24 bovine serum samples.


METHODS
IgG concentration was determined by use of the TIA, and results were compared with those of the radial immunodiffusion (RID) method. Variables were determined, using commercially available reagents and a clinical biochemical analyzer. For the TIA, polyclonal goat anti-bovine IgG (Fc specific) serum, bovine IgG calibrator serum, and polyethylene glycol reaction buffer were used. Sample concentrations were determined by the instrument, using the linear regression method of least squares. The accuracy of this assay was validated by referencing to a purified bovine IgG standard and by recovery of control standards. Parallelism was documented by assay linearity and serial sample dilution linearity. Interference resulting from hemolyzed samples was examined.


RESULTS
The TIA method correlated positively (r = 0.9957) and significantly (P < 0.05) with the RID method, yielding a regression equation with slope of 0.78708 and y-intercept of 1.02102. Bias attributable to hemolysis was not observed.


CONCLUSIONS
The TIA method is automated, accurate, and precise for bovine serum IgG quantification.


CONCLUSIONS
This assay provides sample results in approximately 10 minutes and may be used as an alternative to the manual RID method.
Etzel
L R
LR
AMPC Inc, Ames, IA 50010, USA.
Strohbehn
R E
RE
McVicker
J K
JK
D000818:Animals; D000906:Antibodies / Q000032:analysis / Q000276:immunology; D002417:Cattle / Q000097:blood* / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods / Q000662:veterinary; D006041:Goats; D007118:Immunoassay / Q000379:methods / Q000662:veterinary*; D005779:Immunodiffusion / Q000379:methods / Q000662:veterinary; D007122:Immunoelectrophoresis / Q000379:methods / Q000662:veterinary; D007074:Immunoglobulin G / Q000097:blood* / Q000276:immunology; D007525:Isoelectric Focusing / Q000379:methods / Q000662:veterinary; D009391:Nephelometry and Turbidimetry / Q000379:methods / Q000662:veterinary*; D011092:Polyethylene Glycols; D012044:Regression Analysis; D013997:Time Factors
D000906:Antibodies; D007074:Immunoglobulin G; D011092:Polyethylene Glycols