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Anti-TPO
Anti-Thyroid Peroxidase
REF: DS177708 96 tests
Intended use
Immunoassay for the in vitro quantitative determination of
antibodies to thyroid peroxidase in human serum. The anti‐TPO
determination is used as an aid in the
diagnosis of autoimmune thyroid diseases.
Summary (1, 2, 3, 4, 5, 6)
Thyroid‐specific peroxidase (TPO) is present on the microsomes of
thyrocytes and is expressed at its apical cell surface. In synergy
with thyroglobulin (Tg) this enzyme has an essential function in
the iodination of L‐tyrosine and the chemical coupling of the
resulting mono‐ and di‐iodotyrosine to form the thyroid hormones
T4, T3, and rT3.TPO is a potential autoantigen. Elevated serum
titers of antibodies to TPO are found in several forms of
thyroiditis caused by autoimmunity. The still frequently found term
“microsomal antibody” originates from the time when TPO had not yet
been identified as an antigen in autoimmunity caused by microsomes.
In the
clinical sense the two terms anti‐TPO and microsomal antibody can
be used synonymously; there are differences, however, with regard
to the test methods.
High anti‐TPO titers are found in up to 90 % of patients with
chronic Hashimoto's thyroiditis. In Graves' disease, 70 % of the
patients have an elevated titer.
Although the sensitivity of the procedure can be increased by
simultaneously determining other thyroid antibodies (anti‐Tg,
TSH‐receptor‐antibody - TRAb), a negative finding does not rule out
the possibility of an autoimmune disease. The magnitude of the
antibody titer does not correlate with the clinical activity of the
disease. Initially elevated titers can become negative after
lengthy periods of illness or during remission. If antibodies
reappear following remission, then a relapse is probable.
Whereas the usual microsomal antibody tests employ unpurified
microsomes as an antigen preparation, the anti‐TPO tests use a
purified peroxidase. The two procedures are of comparable
performance in terms of clinical sensitivity, but better lot‐to‐lot
consistency and higher clinical specificity can be expected from
anti‐TPO tests due to the higher quality of the antigen used.The
Enzyme linked immunosorbent assays uses human antigen and rabbit
antihuman IgG antibodies (anti-IgG).
Reagents
Materials provided
• Coated Microplate, 8 x 12 strips, 96 wells. Pre-coated with human
TPO antigen.
• Calibrators, 6 vials, 1 mL each, ready to use; Concentrations:
0(A), 25(B) ,50(C), 100(D), 250(E) and 500(F) IU/mL.
• Enzyme Conjugate, 1 vial, 11.0 mL of HRP (horseradish peroxidase)
labeled rabbit anti-human IgG antibodies (anti-IgG) in Tris-NaCl
buffer containing BSA (bovine serum albumin). Contains 0.1%
ProClin300 preservative.
• Serum Diluent: 1 vial, 11mL. Containing buffer salts and a dye
• Wash Solution Concentrate, 1 vial, 25 ml (40X concentrated),
PBS-Tween wash solution.
• Substrate, 1 vial, 11ml, ready to use, (tetramethylbenzidine)
TMB.
• Stop Solution, 1 vial, 6.0 ml of 1 mol/l sulfuric acid.
• IFU, 1 copy.
• Plate Lid: 2 pieces.
Materials required (but not provided)
• Microplate reader with 450nm and 620nm wavelength absorbent
capability.
• Microplate washer.
• Incubator.
• Plate shaker.
• Micropipettes and multichannel micropipettes delivering 50μl with
a precision of better than 1.5%.
• Absorbent paper.
• Distilled water