Toxoplasma IgG ELISA Kit

Sample

Serum and Plasma

Species Reactivity

Human

Detection Method

ELISA

Intended Use

The Toxoplasma gondii IgG test is quantitative and qualitative immunoassays for the detection of human antibodies in serum or plasma directed against Toxoplasma gondii. The Toxoplasma gondii IgG test allows for the determination of immune status as well as for the detection of intrathecally synthesized antibodies for CSF diagnostics and, by using the corresponding avidity reagent, of IgG antibody avidity determination in order to differentiate acute from past infections.

Contents of Kit

1.ELISA Microplate: 12 × 8
Break apart microtiter test strips each with eight antigen coated single wells.
The coating material is inactivated.
2.Standard Serum (ready-to-use) 2 × 2 ml
Human serum in protein containing phosphate buffer; negative for anti-HIV Ab, HBsAg (Hepatitis B-Virus surface antigen) and anti-HCV Ab;
preservative: < 0.1 % sodium azide; colouring: Amaranth O.
3.Negative Control Serum (ready-to-use) 2 ml
Human serum in protein containing phosphate buffer; negative for anti-HIV Ab, HBsAg (Hepatitis B-Virus surface antigen) and anti-HCV Ab;
preservative: < 0.1 % sodium azide; colouring: Lissamin Green V.
4.Anti-Human IgG Conjugate (ready-to-use) 13 ml
Anti-human IgG polyclonal antibody, conjugated to alkaline phosphatase, stabilised with protein stabilisation solution;
preservative: < 0.1 % methylisothiazolone, < 0.1 % bromnitrodioxane.
5.Washing Solution Concentrate (sufficient for 1000 ml) 33.3 ml
Sodium chloride solution with Tween 20 and 30 mM Tris/HCl, pH 7.4;
preservative: < 0.1 % sodium azide.
6.Dilution Buffer (ready-to-use) 2 × 50 ml
Protein containing phosphate buffer with Tween 20;
preservative: < 0.1 % sodium azide; colouring: 0.01 g/L Bromphenol blue.
7.Stopping Solution (ready-to-use) 15 ml
< 0.1 N sodium hydroxide, 40 mM EDTA.
8.Substrate (ready-to-use) 13 ml
Para-nitrophenylphosphate in solvent free buffer;
preservative: < 0.1 % sodium azide.
9.Quality control certificate with standard curve and evaluation table 2 pages
(quantification of antibodies in IU/ml or U/ml)

Storage

The kit is stored at 2 - 8°C (Avoid Direct Light), and not be frozen or thawed. The product is valid for 12 months. After opening, store at 2 - 8°C, it can be stable for 6 months, avoid contamination.

Performance Characteristics

To determine the performance characteristics of the CD ELISA classic Toxoplasma gondii IgG test, an external comparison study, utilizing sera from 450 patients and healthy individuals, was performed in parallel with an indirect immunofluorescence test. The results indicate a sensitivity of 98.2 % and specificity of 99.4 %.
An internal study was also carried out using sera from 994 pregnant women. An indirect immunofluorescence test was also used as the reference test and discrepant results were further tested in the Sabin-Feldman-Test to determine their true status. The results indicate a sensitivity of 98.2 % and specificity of 99.8 %.

Detection Range

The borderline ranges of the CD ELISA classic Toxoplasma gondii IgG tests are specified on the quality control certificates and indicate the range of borderline test results. Values below this range indicate a negative test result; values above the borderline range are interpreted positive.
The limits of quantification are specified on the quality control certificate of the CD ELISA classic Toxoplasma gondii IgG. The linearity of dilution within this range has been demonstrated in comprehensive evaluation studies. In case a patient sample shows a test result above the upper limit of quantification, the sample may be tested at a higher dilution. The resulting antibody activity must then be multiplied by the additional dilution factor.
5 - 500 IU/mL

General Description

Optimum results can only be achieved if the instructions are strictly followed. Only use ELISA reagents when using ELISA immunoassays. The components must not be exchanged for reagents of other manufacturers. Standard and control sera of ELISA immunoassays are defined exclusively for the test kit to be used and must not be used in other lots. Washing solution, substrate and stop solution can be used for all ELISA immunoassays irrespective of the lot and the test.
Each ELISA test contains a ready-to-use sample dilution buffer. In some cases the use of special dilution buffers is necessary to guarantee consistent quality and reliable results. The dilution buffers can be used irrespective of the lots.
There are three different conjugate concentrations for each immunoglobulin class (IgA, IgG, IgM), indicated on the label as + (low), ++ (medium) and +++ (high). Conjugates with the same concentration and of the same immunoglobulin class are interchangeable and can be used for other ELISA immunoassays irrespective of the lot and the test. Dilution or alteration of the reagents may result in a loss of sensitivity. Use aseptic techniques when removing aliquots from the reagent tubes to avoid contamination.
Reproducibility of test results is dependent on thorough mixing of the reagents. Agitate the flasks containing control sera before use and also all samples after dilution (e.g. by using a vortex mixer).
Be sure to pipette carefully and comply with the given incubation times and temperatures. Significant time differences between pipetting the first and last well of the microtiter plate when dispensing samples and control sera, conjugate or substrate can result in different pre-incubation times, which may influence the precision and reproducibility of the results. Avoid exposure of reagents to strong light during storage and incubation.
Adequate washing avoids test unspecificities. Therefore, the washing procedure should be carried out carefully. All of the flat bottom wells should be filled with equal volumes of washing buffer. At the end of the procedure ensure that the wells are free of all washing buffer in order to avoid uncontrolled dilution effects. Avoid foaming!
Reagents must be tightly closed after use to avoid evaporation and contamination. Take care not to mix-up the caps of the bottles and/or vials.
The ELISA immunoassay is only valid if the lot-specific validation criteria on the quality control certificate are fulfilled.