GoldHealth Medical PPR Competition Antibody Detection ELISA Kits

Product Description

1. Usage
   Designed for the qualitative detection of PPRV antibodies in sheep and goat serum, this kit evaluates the immune status following Peste des petits ruminants vaccination and aids in the serological diagnosis of infected animals.

2. Principle
   Utilizing a competition ELISA method, the kit features PPRV antigen pre-coated on enzyme micro-well strips. During testing, serum samples and Monoclonal Enzyme conjugate are added. If PPRV-specific antibodies are present, they bind to the coated antigen, preventing enzyme-labeled monoclonal antibodies from binding. Conversely, in the absence of specific antibodies, no binding occurs. After washing to remove unbound components, a substrate is added to form a blue product through enzymatic catalysis. The reaction is stopped, and absorbance is measured at 450nm/630nm using a microplate reader.

Kit Components

• PPRV antigen-coated microplate: 96T x 2
• Enzyme conjugate: 12ml
• Sample dilution: 12ml
• PPRV-IgG Negative control serum: 1.5ml
• PPRV-IgG Positive control serum: 1.5ml
• Substrate: 12ml x 2
• Stop solution: 12ml
• 10x concentrated washing buffer: 50ml
• Adhesive Foil: 2 pieces
• Instruction manual: 1 piece

Materials Required but Not Provided

• Microplate Reader (dual wavelength: 450/630 nm)
• Precision micropipettes (single-channel 10-100ul, 0.5-10ul, multi-channel 30-300ul)
• Constant temperature incubator or water bath
• Oscillator
• Disposable tips (10ul, 200ul)
• Deionized water

Sample Requirements

• Samples should be bacteria-free sheep and goat serum. Store at 2-8°C for up to one week; for longer storage, keep at -20°C.
• Avoid samples with severe hemolysis, precipitate, bacterial contamination, or protein suspension.

Preparation

• Allow ELISA reagents to reach room temperature (20-25°C) for 30 minutes before use. Ensure the microplate is dry before opening.
• Washing solution preparation: Dilute the 10x concentrated washing buffer with deionized water at a 1:10 ratio. Crystallization in the concentrated buffer is normal; dissolve at 37°C.

Test Procedure

1. Use the pre-coated microplate, setting 2 wells each for positive and negative controls. Add 50µl of respective control serum. For sample wells, add 40µl sample dilution and 10µl serum sample, followed by 50µl Enzyme conjugate. Mix gently, cover with adhesive foil, and incubate at 37°C for 30 minutes.
2. Discard the liquid, add 300µl diluted washing buffer per well, let stand for 30 seconds, then discard. Repeat 5 times and dry with absorbent paper.
3. Add 100µl substrate solution per well, mix evenly, cover, and incubate at 37°C in the dark for 15 minutes.
4. Add 50µl stop solution per well to halt the reaction and measure results within 10 minutes.

Results Judgement

• Read OD values at 450nm (630nm reference).
• Validity criteria: Negative control OD ≥ 0.60, Positive control OD < 0.40.
• Calculation: Sample OD / Average OD of Negative control = S/N value.
• Interpretation: S/N value ≥ 0.5: Negative, S/N value < 0.5: Positive.

Precautions and Warnings

• Carefully read the manual before use.
• Do not use expired reagents or mix reagents from different lots.
• Dispose of experimental waste via high-pressure steam sterilization or sodium hypochlorite treatment.
• Balance microplates to room temperature before opening; store unused plates and reagents appropriately.
• Use micropipettes for accurate sample and reagent addition.
• Ensure washing buffer fills wells without overflow to avoid cross-contamination.
• Stop solution is corrosive; wash immediately if contact occurs.

Storage and Expiry

• Store at 2-8°C in the dark; do not freeze. Expiry: