Contents of Kit
1. Microtiter plate, 12 breakable microtiter strips (ready-to-use), 8 wells per strip, each well coated with rabbit IgG
2. Calibrator 0 - 4. 5 × 1 mL, Colored dilutions of human serum (ready-to-use; contains ProClin 950). The antibody activities are indicated on the quality control certificate.
3. Negative control. 1 × 1 mL, Colored dilution of human serum (ready-to-use; contains ProClin 950). The antibody activity is indicated on the quality control certificate.
4. Positive control. 1 × 1 mL, Colored dilution of human serum (ready-to-use; contains ProClin 950). The antibody activity is indicated on the quality control certificate.
5. Sample diluent. 1 × 100 mL, Colored solution (ready-to-use; contains ProClin 950)
6. Wash buffer 10×. 1 × 100 mL, Concentrated solution (10×; contains ProClin 950)
7. Conjugate IgM. 1 × 15 mL, Colored solution of polyclonal anti-humanIgM antibody conjugated to horseradish peroxidase (ready-to-use;contains ProClin 950)
8. Substrate TMB. 1 × 15 mL, 3,3´,5,5´-Tetramethylbenzidin (ready-to-use)
9. Stop solution. 1 × 15 mL (75 mL), H2SO4 0.25M.
Storage
Upon receipt, all test components must be stored at 2 °C to 8 °C, preferably in the original kit box. If stored properly in their original containers, all components are stable until their expiry date. All components are stable for at least 2 months after opening when stored properly at 2 °C to 8 °C.
Detection Range
Reliable accuracy, trueness, precision, linearity and recovery of test results have been observed within the measurement range of the assay from the LoQ to the upper calibrator in comprehensive studies. Samples with test results above the upper calibrator should be reported as >max. Samples with test results below the LoQ should be reported as
General Description
Patients suffering from rheumatoid arthritis (RA) exhibit RF, autoantibodies recognizing the Fc part of IgG. RA or chronic polyarthritis has a yet unknown etiology and represents the most frequent rheumatic inflammatory disorder demonstrating a prevalence rate of up to 1%. One of the typical manifestations of RA is symmetric synovialitis of limb joints often accompanied by involvement of the cervical spinal column.
Beside clinical features one of the criteria of the American College of Rheumatology for the diagnosis of RA is the presence of RF. Up to 80 % of RA patients may demonstrate RF. RF can occur years prior to the onset of disease and RF positive apparently healthy individuals bear a 5 - 40 times higher risk to develop RA. However, patients suffering from other autoimmune, infectious or B-cell lymphoproliferative disorders as well as apparently healthy elderly individuals may develop RF.
High concentrations of RF are often associated with a more severe disease comprising a faster destruction of joints. In addition they are found in patients with extra-articular manifestations such as rheumatoid nodules, polyneuropathy, vasculitis or Sicca syndrome.
RF may belong to the IgG, IgM or IgA isotype whereas IgM RF is the most frequent isotype to be determined in RA patients. Extra-articular manifestations seem to be associated with IgA RF. Like RF of the IgM isotype high concentrations of IgG RF seems to appear with patients suffering from more progressive erosions of joints.
Standard Curve
1. Metrological Traceability
The immunoassay is calibrated using the WHO international reference preparation W1066 (CLB 64/1, Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam, conforming 1st British Standard NIBSC 64/2) as a reference. Quantitative results are expressed in IU/mL.
2. Standard Curve
For generation of a standard curve, the optical signals (optical density, OD) of the calibrators are plotted against their antibody activities and correlated by a 4-parameter logistic (4 PL) fit. Antibody activities of unknown samples can be derived directly from their optical signals by use of the generated standard curve.
3. Criteria of Validity
Test runs are only valid if the following criteria of validity are fulfilled:
- OD CAL 0 < CAL 1 < CAL 2 < CAL 3 < CAL 4
- OD CAL 4 > 1.2
- The negative control must be evaluated negative.
- The positive control must be evaluated positive and present an antibody activity within the validity range indicated on the quality control certificate.
If these criteria are not met, the test is not valid and must be repeated.
4. Troubleshooting
In case of an invalid test run, the expiry dates and storage conditions, incubation times and temperatures, and precise calibration of all instruments used should be verified. If no reason for an invalid test run could be identified, please contact the supplier or manufacturer of the product.
Citations
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