Validation for canine serum of 2 commercially available fluoroimmunoassays time-resolved (TR-FIAS) is designed for analysis of cortisol and free thyroxine (FT4) in human serum do. Included is the study of interference by hemolysis, lipemia, and bilirubinemia. By dissociation enhancement lanthanide fluoroimmunoassay kit, intra-assay coefficient of variation (CV) ranged from 6.4% to 8.7% for cortisol and from 5.3% to 9.8% for the FT4; Interassay CVs ranged from 5.8% to 10.8% and from 3.9% to 14.1%, respectively.
Accuracy was evaluated by Mouse Plasma comparing cortisol and FT4 results obtained with TR-FIA and obtained by enzyme-linked immunosorbent assay validated (ELISA) and equilibrium dialysis (ED) assay, respectively. The regression equation obtained y = 0.57x + 1:18 (r2 = 0.90) for cortisol and y = 0.87x + 0.82 (r2 = 0.93) for FT4.
The limit of detection for cortisol and FT4 was 4.84 nmol / L and 2.68 pmol / L, respectively. Results adrenocorticotropin-stimulation and dexamethasone-suppression test similar to that published previously; Also, the dog serum sample dilution series with a high content of cortisol showed that the TR-FIA is immunologically specific.
Serial dilutions of serum samples with high concentrations of FT4 show methodologic bias, reliance on the binding capacity of serum, indicating that the results obtained with this method should be interpreted with caution. Finally, hemolysis and lipemia significantly interfere with cortisol and FT4 measurement, while bilirubinemia not affect the results.
LipL32 recombinant antigen-based single serum dilution ELISA for detection of canine leptospirosis.
An antigen-based single serum dilution of recombinant enzyme-linked immunosorbent assay (ELISA) was developed to measure the activity of specific antibodies in the serum of dogs with leptospirosis. Recombinant antigen developed and used in the assay was specific to pathogenic serovar of Leptospira.
A linear relationship was found Mouse Serum to exist between antibody titers predicted in a single working dilution of 1: 1000 and corresponding serum titer was observed as determined by serial-dilution method standard. Regression analysis was used to determine the standard curve from which the equation can be derived which allows the demonstration of correlation mentioned.
This equation is then used to convert the corrected absorbance reading of a single working dilution directly into the ELISA antibody titers predicted. This test proved to be a sensitive, specific and accurate compared with standard microscopic agglutination test (MAT).
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