Development and validation of a sandwich ELISA for use in measuring concentrations of canine surfactant protein A in serum of dogs
| Autor: | Hideo Akiyoshi, Katsuhito Sone, Shunji Sugii, Mika Aoki, Fumihito Ohashi |
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| Rok vydání: | 2011 |
| Předmět: |
Lung Diseases
Pathology medicine.medical_specialty Enzyme-Linked Immunosorbent Assay Immunoglobulin G Dogs Affinity chromatography medicine Animals Dog Diseases Antiserum Detection limit Chromatography Pulmonary Surfactant-Associated Protein A General Veterinary biology Chemistry General Medicine Pulmonary edema medicine.disease Antibodies Anti-Idiotypic Surfactant protein A biology.protein Rabbits Protein G Antibody Biomarkers |
| Zdroj: | American Journal of Veterinary Research. 72:833-837 |
| ISSN: | 0002-9645 |
| DOI: | 10.2460/ajvr.72.6.833 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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