LSY-30040-2 Ascaris lumbricoidess (A.l) IgM Antibody ELISA Test Kit

Ascaris lumbricoidess (A.l) IgM Antibody ELISA Test Kit

Catalog No. LSY-30040-2

• Principle

This kit use indirect enzyme-linked immunoassay (ELISA) to detect Ascaris lumbricoidess (A.l) IgM antibody in human serum or plasma qualitatively.

• 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) This kit only used to testing human serum or plasma.

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

3) The serum sample can store at 2-8 ℃ for short term (in 3 days), if for long term, it should be kept at -20℃ or lower, avoid repeated freezing and thawing.

5. Test procedure

1) Washing Solution preparation: Dilute 10X Washing solution with distilled water or deionized water at 1:10 (for example: take 100ml 10X Washing solution, add 900ml distilled water or deionized water, mix),mix it evenly to get washing solution.

2) Sample dilute: Dilute serum sample with sample diluent at 1:100 (for example: 5ul serum + 495ul sample diluent), mix it evenly. Positive control and Negative control do not need to dilute, add it directly.

3) Adding samples and controls: for sample wells, only add 100ul sample to each well; 2 wells for blank control, only add 100ul sample diluent to each well; 2 wells for negative control, only add 100ul negative control to each well; 2 wells for Positive control, only add 100ul Positive control to each well. Cover plate with adhesive foil, incubate in dark at 37 ℃ for 30 minutes.

4) Washing plate: discard the liquid of the well, fill full with washing solution to each well(if washing machine, 250ul/well), washing for 3 times, be static for 1 min for each time; at last time, flap to dry with the absorbent paper.

5) Adding Enzyme Conjugate: Except blank control, add Enzyme Conjugate to each well, 50ul/well, Cover plate with adhesive foil, incubate in dark at 37 ℃ for 30 minutes. Discard liquid of the wells and wash 3 times as described in step 4).

6). Adding substrates: add Substrate A, 50ul/well and Substrate B, 50ul/well into all well, mix them evenly. Cover plate with adhesive foil, incubate in dark at 37℃ for 10 minutes.

7). Stop reaction: Add 50ul Stop solution into all wells to stop reaction. Set zero at blank control, read the OD value at ELISA reader 450nm (630nm as a reference).

6. Reference

1) Cut-off Value (COV) = Average OD Value of Negative control X 2.1 (calculate as 0.07 when the OD Value of Negative control < 0.07).

2) Validation judge

Negative control OD value ≤ 0.10 (If more than 0.10, the test is invalid)

Positive control OD value ≥ 0.50 (If less than 0.50, the test is invalid)

7. Results interpretation

Sample OD value ≥ COV: Ascaris lumbricoidess (A.l) IgM antibody positive;

Sample OD value < COV: Ascaris lumbricoidess (A.l) IgM antibody negative.

Limitation: This kit is for research use only, not for clinical diagnosis.

8. Notes

1) 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 2~8 ℃.

2) Before adding reagent, gently shake dropping bottle and mix even the reagent.

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

4) When incubation, it is recommended to use water bath. If no water bath, put at 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.

9. Storage and expire date

Store at 2~8℃ in dark, no frozen, expiry date: 6 months.