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Technical Literature

1.Intended use
The Thyroid Peroxidase IgG test is a rapid ELISA method for the detection of IgG antibodies to human thyroid peroxidase.  It is intended as an aid to diagnosis of autoimmune thyroid disease.  The components of the kit are for in vitro diagnostic use only.

2. Explanation of the Test
Thyroid peroxidase (TPO) is the major enzyme involved in the multiple steps of thyroid hormone synthesis. Autoantibodies to TPO are associated with both Hashimoto's thyroiditis and Graves’ disease.  The presence of anti-TPO autoantibodies unequivocally confirms autoimmune thyroiditis and they are frequently the presenting indication of underlying disease. The detection of anti-TPO autoantibodies is evidence against goitre or hypothyroidism of non-autoimmune nature.

The test is calibrated against the NIBSC standard 66/387.

3. Principle of the test
Diluted serum samples are incubated with TPO immobilised on microtitre wells.  After washing away unbound serum components, rabbit anti-human IgG conjugated to horseradish peroxidase is added to the wells, and this binds to surface-bound antibodies in the second incubation. Unbound conjugate is removed by washing, and a solution containing 3,3’,5,5’-tetramethylbenzidine (TMB) and enzyme substrate is added to trace specific antibody binding. Addition of Stop Solution terminates the reaction and provides the appropriate pH for colour development.  The optical densities of the standards, controls and samples are measured using a microplate reader at 450nm.  Optical density is directly proportional to antibody activity in the sample. 


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Thyroid manager: Chapter 8 - Hashimoto's thyroiditis. Last updated July 2007.