Foot and mouth disease virus (FMDV) Type O Antibody Test (block ELISA) animal diagnostic kits

Foot and mouth disease virus (FMDV) Type O Antibody Test Kit for bovine or goat (block ELISA)

Catalog No. LSY-30031

1. Principle

This kit is based on block ELISA method to detect specific antibody against foot and mouth disease virus (FMDV) Type O Antibody in serum or plasma of bovine and goat qualitatively, for monitoring antibody after FMDV Type O immune.

If the sample contain FMD virus type O antibodies, it would block the FMD virus type O monoclonal antibody HRP conjugate with the FMD virus type O recombinant antigen coated microtiter plate combination. FMD virus package is bound O board monoclonal antibody HRP conjugate and chromogenic substrate reaction can reflect the status of FMD virus type O whether the sample contains antibodies level of protection.

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) This kit can only detect serum or plasma of bovine and goat.

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

3) The serum sample should store at 2-8 ℃ in storage time (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 buffer preparation: dilute 10X Washing buffer with distilled water or deionized water at 1:10 (for example: take 100ml 10X Washing buffer, mix with 900ml distilled water or deionized water), mix evenly.

2. Adding Sample diluent, 50ul/well.

3. Adding samples: for sample wells, add sample 50ul/well; set 2 wells for blank control well, add sample diluent, 50ul/well; set 2 wells for negative control, add negative control, 50ul/well; set 2 wells for positive control, add positive control, 50ul/well; cover with adhesive foil,incubate in dark at 37 ℃ for 60 minutes.

4. Washing plate:discard the liquid of the well, add diluted washing buffer full of the wells (if use ELISA washer, add washing buffer at 250ul/well), wash for 3 times, be static for 1min each time, at last time, flap to dry with the absorbent paper.

5. Adding enzyme conjugate into all wells (except blank well), 100ul/well, cover with adhesive foil and incubate at 37℃ for 60 minutes.Discard the contents of the wells and wash 3 times as described in step 4.

6. Adding substrates: mix Substrate A and Substrate B evenly before usage according to quantity needed, add mixture 100ul/well, cover with adhesive foil,incubate at 37℃ in dark for 15 minutes.

7. Add Stop solution, 50ul/well. Set zero at blank control, read the OD value at ELISA reader 450nm (630nm as a reference).

6. Reference

1) Calculation of PI

PI=average negative control OD value - sample OD value or positive control OD valuex 100%

Average negative control OD value

2) Validation judge

Negative control OD Value ≥ 0.15 (If less than 0. 5, the test is invalid

Positive control PI ≥ 50% (If less than 50%, the test is invalid)

7. Results interpretation

Sample PI ≥ 50%: positive;

Sample PI < 50%: negative.

8. Limitation

This kit is only suitable for bovine, goat serum.

9. 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) Cover plate with adhesive foil when incubate, do not repeat use adhesive foil.

4) Don't mix components between different kits of different lots. Tight the cap after usage, don’t mix using the caps of different bottles.

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.

10. Storage and expire date

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