Human Anti-TPO IgG ELISA Kit

Sample

Serum

Species Reactivity

Human

Detection Method

sELISA

Intended Use

This microplate based ELISA (enzyme-linked immunosorbent assay) kit is intended for the quantitative determination of human anti-TPO autoantibody (IgG) level in serum. The presence of this autoantibody together with clinical findings and other laboratory tests is a useful tool in the aid of diagnosis of autoimmune thyroid disease.

Contents of Kit

1. Streptavidin Coated Microplate
One microplate with 12 x 8 strips (96 wells total) coated with streptavidin. The plate is framed and sealed in a foil Ziploc bag with a desiccant. This reagent should be stored at 2 – 8°C and is stable until the expiration date on the kit box.
2. Biotinylated TPO
One vial containing 10 ml of ready-to-use biotinylated human TPO solution in a stabilized matrix. This reagent should be stored at 2 – 8°C and is stable until the expiration date on the kit box.
3. Anti-hIgG Tracer Antibody
One vial containing 0.6 mL concentrated horseradish peroxidase (HRP) conjugated anti-human IgG tracer antibody in a stabilized protein matrix. This reagent must be diluted with Tracer Antibody Diluent before use. This reagent should be stored at 2 – 8°C and is stable until the expiration date on the kit box.
4. Tracer Antibody Diluent
One vial containing 12 mL ready-to-use buffer. It should be only used for antibody dilution according to the assay procedures. This reagent should be stored at 2 – 8°C and is stable until the expiration date on the kit box.
5. ELISA Wash Concentrate
One bottle contains 30 mL of 30-fold concentrate. Before use the contents must be diluted with 870 mL of distilled water and mix well. Upon dilution this yields a working wash solution containing a surfactant in phosphate-buffered saline with a nonazide preservative. The diluted wash solution should be stored at room temperature and is stable until the expiration date on the kit box.
6. ELISA HRP Substrate
One bottle contains 12 mL of tetramethylbenzidine (TMB) with hydrogen peroxide. This reagent should be stored at 2 – 8°C and is stable until the expiration date on the kit box.
7. ELISA Stop Solution
One bottle contains 12 mL of 0.5 M sulfuric acid. This reagent should be stored at 2 – 8°C or room temperature and is stable until the expiration date on the kit box.
8. Anti-TPO hIgG Calibrators
Five vials each contain anti-TPO IgG type autoantibody in a liquid bovine serum albumin-based matrix with a non-azide preservative. Refer to vials for exact concentration for each calibrator. After the first use, the calibrators should be stored at -20°C or below for long-term storage.
9. Anti-TPO hIgG Controls
Two vials each contain anti-TPO IgG type autoantibody in a liquid bovine serum albumin-based matrix with a non azide preservative. Refer to vials for exact concentration range for each control. After the first use, the calibrators should be stored at -20°C or below for long-term storage.
10. Patient Sample Diluent
Two bottles each contains 30 mL phosphate buffer with protein stabilizers and preservative. The reagent is ready to use. This reagent should be stored at 2 – 8°C and is stable until the expiration date on the kit box.

Storage

This test kit must be stored at 2 – 8°C upon receipt. For the expiration date of the kit refer to the label on the kit box. All components are stable until this expiration date.

Precision

The intra-assay precision is validated by measuring two samples in a single assay with 20 replicate determinations.

The inter-assay precision is validated by measuring two samples in duplicate in 12 individual assays.

Detection Range

0 - 960 U / mL

Sensitivity

The sensitivity of this anti-TPO IgG ELISA Kit as determined by the 95% confidence limit on 20 duplicate determination of zero calibrator is about 1 U/mL.

General Description

It is a routine practice of measuring serum autoantibodies to thyroglobulin (Tg) and microsomal (TPO) for aid in detecting and monitoring autoimmune thyroid disease. Serum anti-TPO autoantibody and anti-Tg autoantibody are found to be well correlating with histological changes in Harshimoto's thyroiditis. Clinically, positive anti-TPO autoantibody is detected in patients with chronic thyroiditis (70-90%), primary hypothyroidism (~60%), thyrotoxicosis (~50%) and thyroid tumors (~17%); however, anti-Tg autoantibody is mainly identified in patients with Harshimoto's thyroiditis and Graves' disease (40-70%).
Although ELISA technology has applied to detecting these autoantibodies, the high background in normal population would decrease the clinical diagnostic sensitivity and specificity. This high sensitive anti-TPO IgG ELISA kit was developed with proprietary technology that leads to a very low reaction background in normal population and thus would increase the clinical diagnostic sensitivity and specificity.

Standard Curve

A typical absorbance data and the resulting calibrator curve from human anti-TPO IgG ELISA are represented. This curve should not be used in lieu of calibrator curve run with each assay.