Anti-dsDNA IgG test is used for initial diagnosis of Systemic Lupus Erythematosus (SLE) and for diagnosis of SLE different diseases. Besides the determination of high titers of antinuclear antibody (ANA), the determination of autoantibodies against dsDNA is one of the ACR criteria (American College of Rheumatology) for the diagnosis of Systemic Lupus Erythematosus (SLE). The determination of the concentration of antibodies can be used to monitor treatment success and predict possible attacks of the disease (SLE)
Intended Use:
Anti dsDNA IgG kit is an indirect solid phase enzyme immunometric assay (ELISA) designed for the quantitative measurement of IgG class antibodies directed against dsDNA in human serum or plasma. Anti dsDNA IgG is intended for laboratory use only
Specificity:
Comparison test against a commercial reference kit, performed on 36 sera (15 of them positive sera and 21 negative sera) showed a 95.24% specificity.
Sensitivity:
Comparison test against a commercial reference kit, performed on 36 sera (15 of them positive sera and 21 negative sera) showed a 93.33% sensibility.
Test Principle:
Anti-dsDNA IgG test is based on the binding of serum or plasma IgG antibodies on dsDNA coated on the microplates. The antibodies in calibrators, controls or prediluted patient samples bind into the inner surface of the wells. After a 30 minutes incubation the microplate is washed with wash buffer for removing non-reactive serum components. An anti-human-IgG horseradish peroxidase conjugate solution recognizes the IgG class antibodies bound to the immobilized dsDNA antigens. After a 30 minutes incubation any excess of enzyme conjugate, which is not specifically bound, is washed away with wash buffer. A chromogenic substrate solution containing TMB is dispensed into the wells. After 15 minutes of incubation the color development is stopped by adding the stop solution. The solution color changes into yellow. The amount of color is directly proportional to the concentration of the anti-dsDNA IgG antibodies present in the original sample. The concentration of the anti dsDNA IgG antibodies in the sample are calculated through a calibration curve.
Kit Components:
1. dsDNA coated wells, 12x8-well snap-off stips coated with dsDNA.
2. Stop SolutionL 1x15ml
3. Conjugate, 1x15ml
4. Sample diluent, 1x100ml
5. Wash solution, 1x50ml
6. dsDNA standards, 5x1.2ml
Standard 0: 0AU/ml
Standard 1: 15AU/ml
Standard 2: 30AU/ml
Standard 3: 60AU/ml
Standard 4: 240AU/ml
7. Negative control, 1x1.2ml
8. Positive control, 1x1.2ml
Storage and Stability:
Store components at 4 degrees C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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