Medical Elisa Test Kit CE , Toxo IgG Rapid Elisa Test Quantitative Determination

Toxoplasma gondii (Toxo) IgG ELISA test, high accuracy, Elisa Sandwich method, for quantitative measurement

Product Name:  Toxoplasma gondii (Toxo) IgG ELISA test

Intended Use:

Toxoplasma gondii (Toxo) IgG Enzyme-Linked Immunosorbent Assay (ELISA) is intended for the detection and quantitative determination of IgG antibody to Toxoplasma gondii in human sera. This product is not FDA cleared (approved) for use in testing (ie, screening) blood or plasma donors. For in vitro diagnostic Use. High complexity test.

Summary:

Toxoplasma gondii causes toxoplasmosis, a common disease that affects 30-50 of every 100 people in North America by the time they are adults. The mean source of infection is direct contact with cat feces or from eating undercooked meats. Toxoplasmosis generally presents with mild symptoms in immunocompetent individuals; in the immunocompromised patient, however, the infection can have serious consequences. Acute toxoplasmosis in pregnant women can result in result in miscarriage, poor growth, early delivery or stillbirth. Treatment of an infected pregnant woman may prevent or lessen the disease in her unborn child. Treatment of an infected infant will also lessen the severity of the disease as the child grows. IgG and IgM antibodies to Toxoplasma can be detected with 2-3 weeks after exposure. IgG remains positive, but the antibody level drops overtime. ELISA can detect Toxoplasma IgM antibody after one year after infection in over 50% of patients. Therefore, IgM positive results should be evaluated further with one or two follow up samples if primary infection is suspected.

Principle of Test:

Toxoplasma gondii IgG ELISA Kit is a solid-phase immunoanalytical test. The purified, homogeneous antigen

is fixed to each well of the microtiterstrips. Specific antibodies present in the patient’s sample are bound during

the first incubation step. After removing unbound material by washing, the presence of the specific antibodies

is detected using anti-human IgG conjugate during the second incubation. The unbound peroxidase conjugate

is then removed and TMB substrate is added, resulting in the development of a blue colour. The enzyme

reaction is terminated by addition of the stop solution. The intensity of the colour is proportional to the

concentration of the antibodies in the sample.

Kit Components

ASSAY PROCEDURE

1.  Allow the vacuum-closed aluminium bag with strips to reach room temperature. Withdraw an adequate number of strips and put the unused strips into the provided bag and seal it carefully with the desiccant kept inside.
2.  Pipette 100 µL of Sample diluent, Controls and serum samples to the wells according to the pipetting scheme in Plate Layout: fill the first well with Dilution buffer (DIL) to determine the reaction background. Fill the next two wells with Calibrator 1 (CAL 1). The next wells fill with Calibrator 2-4 (CAL 2-4) and Negative control (CONTROL 1). The remaining wells fill with diluted serum samples (S1...). It is satisfactory to apply one serum into one well (S1, S2, S3...). However, if you want to minimize a laboratory error, apply controls and samples as doublets. Cover the strips with the Cover membrane or cover.
3.  Incubate 60 minutes (+/- 5 min.) at 37°C in a humid chamber.
4.  Aspirate the liquid from wells into a collecting bottle containing appropriate disinfectant (see Safety Precautions). Wash and aspirate the wells four times with 300 µl/well of Wash buffer. Avoid cross-contamination between wells! If some liquid remains in the wells, invert the plate and tap it on an adsorbent paper to remove the last remaining drops.
5.  Add 100 µL Peroxidase conjugate r.t.u. into each well. Incubate 30 minutes (+/- 2 min) at 37°C in a humid chamber.
6.   Aspirate and wash four times with 300 µl/well of Wash buffer.
7.  Dispense 100 µl of TMB into each well. Incubate 15 minutes (+/- 30 seconds) at room temperature. The time measurement must be started at the beginning of TMB dispensing. Cover the strips with opaque cover or keep them in the dark during the incubation with TMB.
8.  Stop the reaction by adding 100 µL of Stop solution. Use the same pipetting rhythm as with the TMB to ensure the same reaction time in all wells. Tap gently the microplate for a few times to ensure complete mixing of the reagents.
9.  Read the absorbance at 450 nm with a microplate reader within 10 minutes. It is recommended to use a reference reading at 630 (620) nm.