Quantification of serum total IgE concentration in dogs by use of an enzyme-linked immunosorbent assay containing monoclonal murine anti-canine IgE.
A method for quantification of serum total IgE concentration in dogs by use of an ELISA containing monoclonal mouse anti-canine IgE was developed. Microtitration plates were coated with monoclonal mouse anti-canine IgE. Test sera and reference serum dilutions were added, followed by biotinylated monoclonal mouse anti-canine IgE. Avidin-alkaline phosphatase conjugate was added, and color development was measured spectrophotometrically, using a microtitration plate reader. Quantitative results were obtained by assigning to a reference serum a value of 100 IgE units/ml. Absorbance values of unknown samples were converted into IgE units by comparison with a standard curve generated by measurement of reference serum dilutions. Intra- and interassay coefficients of variation were 5 and 7%, respectively, and assay sensitivity was 1 U/ml. The assay was used to establish a normal range for total IgE concentrations in 30 healthy dogs. Total IgE concentration in healthy dogs followed a skewed distribution and ranged from < 1 to 91.2 U/ml, with a geometric mean value of 7.1 U/ml. The IgE concentration was remarkably stable in serum samples subjected to 25 freeze/thaw cycles or incubation at approximately 25 C (room temperature) for up to 10 days. Comparison of total IgE concentrations in 23 serum samples assayed by use of double-overlay radial immunodiffusion and ELISA yielded correlation coefficient of 0.94. Comparison of the reference serum standard curve with serial dilutions of a purified IgE solution of known concentration yielded a range of values for the IgE unit of 0.7 to 2.0 micrograms.
Department of Medical Sciences, School of Veterinary Medicine, University of Wisconsin-Madison 53706.
P B
PB
Hill
D J
DJ
DeBoer
D000818:Animals; D000911:Antibodies, Monoclonal; D004184:Dirofilariasis / Q000097:blood / Q000276:immunology; D004283:Dog Diseases; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary; D005260:Female; D005779:Immunodiffusion / Q000379:methods; D007073:Immunoglobulin E / Q000097:blood*; D008297:Male; D010272:Parasitic Diseases / Q000097:blood / Q000276:immunology; D010273:Parasitic Diseases, Animal; D012016:Reference Values; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D007073:Immunoglobulin E