Evelyn Skopek1, Martina Patzl, Raymond F Nachreiner. 1. Department of Natural Science, Institute of Biochemistry, University of Veterinary Medicine, Veterinärplatz 1, 1210 Vienna, Austria.
Abstract
OBJECTIVE: To establish a sensitive test for the detection of autoantibodies against thyroid peroxidase (TPO) in canine serum samples. SAMPLE POPULATION: 365 serum samples from dogs with hypothyroidism as determined on the basis of serum concentrations of total and free triiodothyronine (T3), total and free thyroxine (T4), and thyroid-stimulating hormone, of which 195 (53%) had positive results for at least 1 of 3 thyroid autoantibodies (against thyroglobulin [Tg], T4, or T3) and serum samples from 28 healthy dogs (control samples). PROCEDURE: TPO was purified from canine thyroid glands by extraction with detergents, ultracentrifugation, and precipitation with ammonium sulfate. Screening for anti-TPO autoantibodies in canine sera was performed by use of an immunoblot assay. Thyroid extract containing TPO was separated electrophoretically, blotted, and probed with canine sera. Alkaline phosphatase-conjugated rabbit anti-dog IgG was used for detection of bound antibodies. RESULTS: TPO bands were observed at 110, 100, and 40 kd. Anti-TPO autoantibodies against the 40-kd fragment were detected in 33 (17%) sera of dogs with positive results for anti-Tg, anti-T4, or anti-T3 autoantibodies but not in sera of hypothyroid dogs without these autoantibodies or in sera of healthy dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The immunoblot assay was a sensitive and specific method for the detection of autoantibodies because it also provided information about the antigen. Anti-TPO autoantibodies were clearly detected in a fraction of hypothyroid dogs. The value of anti-TPO autoantibodies for use in early diagnosis of animals with thyroid gland diseases should be evaluated in additional studies.
OBJECTIVE: To establish a sensitive test for the detection of autoantibodies against thyroid peroxidase (TPO) in canine serum samples. SAMPLE POPULATION: 365 serum samples from dogs with hypothyroidism as determined on the basis of serum concentrations of total and free triiodothyronine (T3), total and free thyroxine (T4), and thyroid-stimulating hormone, of which 195 (53%) had positive results for at least 1 of 3 thyroid autoantibodies (against thyroglobulin [Tg], T4, or T3) and serum samples from 28 healthy dogs (control samples). PROCEDURE: TPO was purified from canine thyroid glands by extraction with detergents, ultracentrifugation, and precipitation with ammonium sulfate. Screening for anti-TPO autoantibodies in canine sera was performed by use of an immunoblot assay. Thyroid extract containing TPO was separated electrophoretically, blotted, and probed with canine sera. Alkaline phosphatase-conjugated rabbit anti-dog IgG was used for detection of bound antibodies. RESULTS:TPO bands were observed at 110, 100, and 40 kd. Anti-TPO autoantibodies against the 40-kd fragment were detected in 33 (17%) sera of dogs with positive results for anti-Tg, anti-T4, or anti-T3 autoantibodies but not in sera of hypothyroiddogs without these autoantibodies or in sera of healthy dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The immunoblot assay was a sensitive and specific method for the detection of autoantibodies because it also provided information about the antigen. Anti-TPO autoantibodies were clearly detected in a fraction of hypothyroiddogs. The value of anti-TPO autoantibodies for use in early diagnosis of animals with thyroid gland diseases should be evaluated in additional studies.
Authors: Alisdair M Boag; Michael R Christie; Kerry A McLaughlin; Harriet M Syme; Peter Graham; Brian Catchpole Journal: PLoS One Date: 2015-11-30 Impact factor: 3.240
Authors: Julia D Labadie; Sheryl Magzamen; Paul S Morley; G Brooke Anderson; Janna Yoshimoto; Anne C Avery Journal: J Vet Intern Med Date: 2019-01-21 Impact factor: 3.333