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  • Author or Editor: Shunji Sugii x
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Sandwich elisa were developed to quantitatively determine conglutinin (cg), mannan-binding protein (mbp), and serum amyloid-P component (sap) in the sera of cattle. The elisa system was found to have high repeatability for quantitation of these serum proteins at concentration as low as 5 ng/ml. From results obtained for 10 healthy cows aged 2 to 7 years, mean ± sd serum concentrations were 56.5 ± 14.4 μg of cg/ml, 2.37 ± 0.87 μg of mbp/ml, and 11.14 ± 3.92 μg of sap/ml, respectively. Values in 6 healthy heifer calves aged 6 months were 3.45 ± 1.22 μg/ml for cg, 1.71 ± 0.96 μg/ml for mbp, and 5.45 ± 2.75 μg/ml for sap, respectively. Concentrations in 9 healthy bullocks aged 6 months were 1.83 ± 0.66 μg/ml for cg, 1.04 ± 0.63 μg/ml for mbp, and 4.9 ± 1.13 μg/ml for sap, respectively.

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in American Journal of Veterinary Research


Objective—To develop and evaluate a sandwich ELISA incorporating rabbit antiserum specific for canine surfactant protein A (SP-A) for use in measuring concentrations of SP-A in serum of dogs.

Sample—Serum samples obtained from 6 healthy dogs and 3 dogs with pulmonary disease.

Procedures—Rabbit antiserum was prepared against purified canine SP-A. The IgG fraction was isolated via protein G affinity chromatography and was then biotinylated. The sandwich ELISA was performed by use of anti-SP-A antibody (IgG) preabsorbed with sera from healthy dogs. Validity of the ELISA was confirmed by determination of the detection limit, precision, reproducibility, and accuracy. Serum SP-A concentrations were measured in 6 healthy dogs and 3 dogs with pulmonary disease.

Results—Detection limit of the ELISA was 2.0 ng/mL. Within- and between-assay coefficients of variation ranged from 3.8% to 14.1% and from 15.5% to 35.6%, respectively. The observed-to-expected recovery ratio ranged from 77.1% to 89.9%. Serum SP-A concentrations measured by use of the ELISA were ≤ 2.3 ng/mL in the 6 healthy dogs, 25.6 ng/mL in a dog with severe cardiac pulmonary edema, 8.3 ng/mL in a dog with pneumonia, and 10.1 ng/mL in a dog with lung lobe torsion.

Conclusions and Clinical Relevance—The sandwich ELISA was found to be useful for measuring purified canine SP-A concentrations and canine SP-A concentrations in serum samples. The ELISA was precise, reproducible, and accurate. The ELISA may be beneficial in assessing serum concentrations of canine SP-A as a potential biomarker of pulmonary diseases in dogs.

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in American Journal of Veterinary Research


Objective—To determine whether cross-reactivity exists between canine chromogranin A (CgA) and anti-human CgA antibody and investigate the usefulness of plasma CgA concentration measurements as an index of acute stress responses in dogs.

Animals—12 healthy Beagles.

Procedure—Canine CgA was extracted and purified from canine adrenal glands of cadaver dogs for studying cross-reactivity with anti-human CgA antibody. Western blotting with anti-human CgA antibody was performed. Blood samples were collected from dogs at 0, 10, 20, 30, 40, 60, 120, and 180 minutes after IV administration of saline (0.9% NaCl) solution or insulin. Canine plasma CgA concentrations were determined by use of a CgA ELISA kit with rabbit antiserum against the carboxy-terminal fragment of human CgA. Plasma cortisol and catecholamine (ie, norepinephrine and epinephrine) concentrations were measured by use of an ELISA and a high-performance liquid chromatography method, respectively.

Results—Purified canine CgA was specifically detected by use of western blot analysis and an ELISA with anti-human CgA antibody. An increase in plasma CgA concentrations was observed in insulin-induced hypoglycemic dogs. Changes in plasma CgA concentration were correlated with changes in plasma cortisol or catecholamine concentrations of hypoglycemic dogs.

Conclusions and Clinical Relevance—Use of the CgA ELISA kit for determination of human plasma CgA concentrations is applicable to the measurement of canine plasma CgA concentrations. Canine plasma CgA concentrations, along with measurements of plasma cortisol and catecholamine concentrations, correctly reflect insulin-induced hypoglycemic stressed conditions in dogs. Measurement of canine plasma CgA concentrations may provide a useful index for evaluation of an acute stress response. (Am J Vet Res 2005;66:1830–1835)

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in American Journal of Veterinary Research