CDSimple™ T4, T3, TSH Chemiluminescent ELISA Kit

Sample

Serum, plasma

Species Reactivity

Human

Intended Use

The Quantitative Determination of Free Thyroxine; Free Triiodothyronine; Thyrotropin Concentration for a comprehensive thyroid status of a Human Serum or Plasma sample by a Microplate Enzyme Immunoassay, Colorimetric.

Contents of Kit

Reagents for 2 X 96 well Microplate, provided
A. CDSample™ Free Thyroid Calibrators - 1ml/vial- A-F
Six (6) vials of Thyroid CDSample™ human serum calibrators dispensed in vials with the concentrations as listed in the Table. Store at 2-8°C. A preservative has been added.
*Exact levels are given on the labels on a lot specific basis*

B. Strept fT4 Enzyme Reagent - 7 ml/vial
One (1) vial of thyroxine-horseradish peroxidase (HRP) conjugate in a bovine albumin-stabilizing matrix. A preservative has been added. Store at 2-8°C.
C. Strept fT3 Enzyme Reagent - 7 ml/vial
One (1) vial of triiodothyronine -horseradish peroxidase (HRP) conjugate in a bovine albumin-stabilizing matrix. A preservative has been added. Store at 2-8°C.
D. TSH Enzyme Reagent — 13ml/vial
One (1) vial containing enzyme labeled affinity purified polyclonal goat antibody, biotinylated monoclonal mouse IgG in buffer, dye, and preservative. Store at 2-8°C.
E. Strept fT4 Biotin Reagent - 7ml/vial
One (1) vial of biotinylated anti-thyroxine (sheep) reagent in a protein-stabilized matrix. A preservative has been added. Store at 2-8°C.
F. Strept fT3 Biotin Reagent - 7ml/vial
One (1) vial of biotinylated anti-triiothyronine (sheep) reagent in a protein-stabilized matrix. A preservative has been added. Store at 2-8°C.
G. Streptavidin Coated Microplate - 2 x 96 wells
Two 96-well microplates coated with streptavidin and packaged in an aluminum bag with a drying agent. Store at 2-8°C.
H. Wash Solution Concentrate - 20ml
One (1) vial containing a surfactant in buffered saline. A preservative has been added. Store at 2-8°C.
I. Substrate Reagent - 2 x 12ml/vial
Two (2) amber bottles contain tetramethylbenzidine (TMB) and hydrogen peroxide (H₂O₂ ) in buffer. Store at 2-8°C.
J. Stop Solution - 2 x 8ml/vial
One (1) vial contains a strong acid (0.5M H₂SO₄ ). Store at 2-8°C.
K. Product Insert.
Note 1: TSH concentrations were calibrated using a reference preparation, which was
assayed against the WHO 2^nd, IRP 80/558
Note 2: Do not use reagents beyond the kit expiration date.
Note 3: Avoid extended exposure to heat and light. Opened reagents are stable for sixty (60) days when stored at 2-8°C. Kit and component stability are identified on the label.

Storage

Store at 2°C to 8°C

Precision

The within and between assay precision of the Free Thyroid Panel were determined by analvses on three different levele of nool control sera The number. mean values. standard deviation and coefficient of variation for each of these control sera are presented in Table 4 and Table 5 (fT4), Table 6 and Table 7 (fT3) and Table 8 and Table 9 (TSH).

Sensitivity

The fT4 procedure has a sensitivity of 0.04 ng/dl. The sensitivity was ascertained by determining the variability of the 0 ng/dl serum calibrator and using the 2ơ (95% certainty) statistic to calculate the minimum dose.
The fT3 procedure has a sensitivity of 0.04 pg/mL. The sensitivity was ascertained by determining the variability of the 0 pg/mL serum calibrator and usino the 2ơ (95% certainty) statistic to calculate the minimum dose.
The TSH sensitivity (detection limit) was ascertained by determining the variability of the 0 ulU/mL serum calibrator and using the 2ơ (95% certainty) statistic to calculate the minimum dose: For 1 hr incubation =0.065 ulU/mL.

General Description

Measurements of thyroid hormones (fT3, fT4 and TSH) are generally regarded as invaluable in-vitro diagnostic tests for assessing thyroid function.
The Combination Free Thyroid Panel provides the convenience of combination calibrators, universal plate and flexible reagent selection allowing technicians to perform a variety of assay designs. In this method, serum reference, patient specimen, or control is first added to a microplate well. Enzyme-fT4 (fT3) conjugate and biotinylated fT4 or fT3 antibody are added, and the reactants are mixed. In the case of TSH, the biotinylated and enzyme conjugate are added in one step. A reaction results between the enzyme conjugate, biotinylated conjugate and the native thyroid hormone (fT3, fT4 or TSH) for the antibody combining sites. Immobilization takes place through the reaction of the incorporated biotin and streptavidin coated on the well. After the completion of the required incubation period, the bound enzyme conjugate is separated from the unbound enzyme conjugate by aspiration or decantation. The activity of the enzyme present on the surface of the well is quantitated by reaction with a suitable substrate to produce color.
The employment of several serum references of known thyroid hormone concentration(s) permits construction of a graph of activity and concentration. From comparison to the dose response curve(s), an unknown specimen's activity can be correlated with hormone concentration.