Human Brucella IgG ELISA Kit

Sample

Serum, plasma

Species Reactivity

Human

Intended Use

The ELISA test kit provides a semiquantitative or quantitative in vitro assay for human antibodies of the immunoglobulin class IgG class against Brucella abortus in serum or plasma for the diagnosis of brucellosis.

Contents of Kit

1. Microplate wells coated with antigens, 12 microplate strips each containing 8 individual break-off wells in a frame, ready for use 12 x 8 STRIPS
2. Calibrator 1, 200 RU/ml (IgG, human), ready for use, 1 x 2.0 ml CAL1
3. Calibrator 2, 20 RU/ml (IgG, human), ready for use, 1 x 2.0 ml CAL2
4. Calibrator 3, 2 RU/ml, (IgG, human), ready for use, 1 x 2.0 ml CAL3
5. Positive control, (IgG, human), ready for use, 1 x 2.0 ml POS CONTROL
6. Negative control, (IgG, human), ready for use, 1 x 2.0 ml NEG CONTROL
7. Enzyme conjugate, peroxidase-labelled anti-human IgG (rabbit), ready for use, 1 x 12 ml CONJUGATE
8. Sample buffer, ready for use, 1 x 100 ml SAMPLE BUFFER
9. Wash buffer, 10x concentrate, 1 x 100 ml WASH BUFFER 10x
10. Chromogen/substrate solution, TMB/H₂O₂, ready for use, 1 x 12 ml SUBSTRATE
11. Stop solution, 0.5 M sulphuric acid, ready for use, 1 x 12 ml STOP SOLUTION
12. Test instruction --- 1 booklet
13. Quality control certificate --- 1 protocol

Storage

The test kit has to be stored at a temperature between +2°C to +8°C. Do not freeze. Unopened, all test kit components are stable until the indicated expiry date.

Performance Characteristics

Calibration: As no quantificated international reference serum exists for antibodies against Brucella abortus, the calibration is performed in relative units (RU).
For every group of tests performed, the extinction readings of the calibrators and the relative units and/or ratios determined for the positive and negative controls must lie within the limits stated for the relevant test kit lot. A quality control certificate contain
ing these reference values is included. If the values specified for the controls are not achieved, the test results may be inaccurate and the test should be repeated. The binding activity of the antibodies and the activity of the enzyme used are temperature-dependent. It is therefore recommended using a thermostat in all three incubation steps. The higher the room temperature (+18°C to +25°C) during the incubation steps, the greater will be the extinction. Corresponding variations apply also to the incubation times. However, the calibrators subject to the same influences, with the result that such variations will be largely compensated in the calculation of the result.
Antigen: The antigen used is Brucella abortus of strain W99.

Detection Limit

The lower detection limit is defined as the mean value of an analyte-free sample plus three times the standard deviation and is the smallest detectable antibody titer. The lower detection limit of the Anti-Brucella abortus ELISA (IgG) is 0.4 RU/ml.

General Description

The Anti-Brucella abortus ELISA is a supplement to the direct pathogen detection for the diagnosis of Brucellosis. Taking into consideration the clinical symptoms, detection of IgM and/or IgA antibodies and a clear titer increase of IgG antibodies in a serum pair taken after at least 2 weeks time can be seen as indication for an acute Brucella infection. Moreover, serological investigations are suitable for clarification of epidemiological questions.

Standard Curve

The following plot is an example of a typical calibration curve. Please do not use this curve for the determination of antibody concentrations in patient samples.