Canine/ Feline/ Rabbit/ Rat Toxoplasma gondii IgG antibody ELISA kit TOXO Ab

Canine/ Feline/ Rabbit/ Rat Toxoplasma gondii IgG antibody ELISA kit

Catalog No. LSY-30032

1. Principle

Toxoplasmosis human zoonotic parasites, its pathogen Toxoplasma (Toxoplasma gondii, TO), by the parasite of cats, dogs and other animals to people and cause toxoplasmosis people, often causing damage to the eyes, the brain, infected pregnant women may also cause teratogenic even abortion. Detection of Toxoplasma gondii antibodies can aid the diagnosis of infection.

This kit can be used to detect the cat, dog, rabbit, mouse serum specific toxoplasma antibody (IgG).

1. Reagents and contents

3.Material required not provided

1) Microplate Reader with 450nm and 630nm

2) 37 ℃thermostatic device

3) Micropipettes, adjustable.

4. Sample requirement

1) To get the best test result, avoid using sample with severe hemolysis, precipitate, contaminated by bacteria or protein suspension.

2) The serum sample storage time should be less than 1 week at 2-8 ℃, if for long term, it should be kept at -20℃ or lower.

5. Test procedure

1. Sample diluent: Dilute serum with Sample diluent (No.5 solution) at 1:100. Positive control and negative control do not need dilute, add directly;

2. Adding samples and controls: Add 100μl diluted serum sample into all sample well. Set 1 well for negative control, add 100ul negative control; 1 well for positive control, add 100ul positive control; 1 well for blank control, add 100ul Sample diluent (No.5 solution).Incubate in dark at 37 ℃ for 30 minutes,then discard the liquid of the well, add 1 drop washing buffer (No.2 solution) into each well and fill with distilled water to full, incubate for 30 seconds, then discard the liquid, continue perform another 4 washing cycles as above, incubate for 30 seconds each time, then discard the liquid, flap to dry with the absorbent paper.

4. Adding Enzyme conjugate: Add 2 drops of Enzyme Conjugate (No.1 solution) into all wells (except blank well) andincubate at 37℃ for 30 minutes.Discard the contents of the wells and wash 5 times as described in step 3.

5. Adding substrates: Add 1 drop Substrate A (No.3 solution) and 1 drop Substrate B (No.4 solution) into all well,incubate at 37℃ for 10 minutes.

6. Add one drop Stop solution (No.6 solution) into all wells to stop reaction (blue color become yellow). Set zero at blank control, read the OD value at ELISA reader 450nm (630nm as a reference).

6. Results interpretation

1)Judge by eye: the negative control should be near colorless, positive control should be yellow, it means the test is valid. The sample well is near colorless, it means Negative, the sample well is yellow, it means Positive.

2) Judge by ELISA Reader:

Set zero at blank control, read the OD value at ELISA reader 450nm (630nm as a reference).

Sample OD value ≥ 2.1 * Negative control OD Value, the result is Positive;

Sample OD value < 2.1 * Negative control OD Value, the result is Negative;

When the Negative control OD Value is less than 0.07, calculate as 0.07.

7. Notes

1) TOX-Ag coated plates should be sealed and moisture-proof. MicroWell plate removed from the refrigerated environment should be balanced moisture to dry at room temperature, then can be opened. Put back unused MicroWell plate into dry foil bag and sealed at 4 ℃.

2) Before dropping reagent, gently shake dropping bottle and mix even the reagent, discard the first drop; Make sure the dropping bottle is vertical when dropping.

3) Screw down the cap after use, don't mix caps between different bottles and don't mix components between different kits

4) When incubate at 37℃, suggested to use water bath. If there is no water bath, put wet box with adhesive foil in 37℃ incubator.

5) The test kit and Any samples should be regarded as the source of infection to properly handle. Stop solution is corrosive, be careful to use.

8. Storage and expire date

Store at 2~8℃ in dark, no frozen, expiry date: 12 months.