LSY-30012 Canine rabies virus antibody ELISA Assay Kit 1 plate or 2 plates per kit

Green Spring Canine rabies virus antibody ELISA kit

Catalog No. LSY-30012

1. Principle

This kit use enzyme-linked immunoassay (ELISA) to detect specific antibody against Canine rabies virus in dog serum or plasma, for monitoring antibody after dog immune and aided inspection of this disease.

2. 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 dog 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 and Cut-off control dilute: Dilute serum sample and 20X Cut-off control with sample diluent at 1:20 (for example: 10ul serum + 190ul sample diluent), mix it evenly. 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 Cut-off control, only add 100ul Cut-off 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 using 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, 100ul/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: according to the quantity needed, take same volume of Substrate A and Substrate B, mix them evenly before use, then add into all well,100ul/well, Cover plate with adhesive foil, incubate in dark at 37℃ for 15 minutes.

7). Stop reaction: Add 50ul Stop reaction 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

Validation judge

Negative control OD average value ≤ 0.15 (If more than 0.15, the test is invalid)

  Cut-off control OD average value ≥ 0.30 (If less than 0.30, the test is invalid)

7. Results interpretation

Sample OD value ≥ cut-off control OD average value: RBV-Ab positive, and it means the antibody level higher than 0.50 IU/ ml;

Sample OD value < cut-off control OD average value: RBV-Ab negative, and it means the antibody level lower than 0.50 IU/ml.

Limitation: This kit is for screening only, test results should be confirmed in accordance with the relevant standards.

8. Notes

1) The cut-off control is 20X concentrated solution, dilute at 1:20 firstly, then use.

2) 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 ℃.

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

  4) When incubation, must cover the plate with adhesive foil, do not repeat use the adhesive foil.

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

6) 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: 12 months.

Shenzhen Lvshiyuan Biotechnology Co., Ltd

D Building, National Biological Industrial Park of Marinelife, No.2 Binhai Road, Dapeng, Shenzhen, 518120 China

Tel. 86-755-28438788

Fax 86-755-28938800

Email:   info@lsybt.com

www.lsybt.com