Porcine Parvovirus Antibody ELISA Test Kit for Pig 192 Wells/Kit GMP

1. Porcine Test Kit Usage

GreenSpring® Porcine Parvovirus (PPV) Antibody ELISA Kit is used for qualitative detection of Porcine Parvovirus (PPV) IgG antibody in swine serum; evaluation of porcine parvovirus immune conditions in pig farms and epidemiological investigation of Porcine Parvovirus.

2. Principle

GreenSpring® Porcine Parvovirus (PPV) Antibody ELISA Detection Kit is made from antigen-coated microtiter plates (PPV antigen-coated) and other reagents. It applies the principle of solid-phase ELISA to PPV-Ab in serum, and then adds enzyme conjugate to the microplate to specifically bind to the complex of coated antigen + PPV-Ab + enzyme-labeled anti-pig IgG antibody. With TMB substrate, it produces a certain amount of color. The depth of color is related to the content of PPV-Ab. When the color value is greater than the critical value, the pigs are well inoculated or have natural infection.

3. Porcine Test Kit components

4. Materials Required But Not Provided

1) Microplate Reader (double-wave length: 450/630 nm).

2) Precise micropipette (single-channel 1-100ul,0.5-10ul,multi-channel 30-300ul)

3) Constant temperature box or water bath box.

4) Oscillator.

5) Disposable tips (10ul, 200ul)

6) Deionized water

5. Sample requirement

1) The samples are porcine serum, which should be collected with no bacteria. The storage time should be less than 1 week at 2-8 ℃, if for long term, it should be kept at -20℃.

2) Avoid to use the samples with severe hemolysis, precipitate, contaminated by bacteria or protein suspension.

6. Preparation

1) Bring ELISA reagents to the room temperature (20-25 ℃) for 30 min to get best results. Microplate should return to room temperature and dry before open package.

2) Sample dilute: Dilute sample with the sample diluent at 40 times.(5ul serum + 195ul sample diluent), the diluted sample need to mix evenly to get better results.

3) Washing solution preparation: Dilute the 20×concentrated washing buffer with deionized water at 20 times. (50ml 20×concentrated washing buffer + 950ml deionized water ) It is normal if there is crystallization in the 20×concentrated washing buffer, put at 37℃ until completely dissolved.

7. Procedure

1) Take out the coated plates (Can be detached) and record the sample position on a worksheet. Set 2 wells for negative control serum, add undiluted negative control serum, 2 wells for positive control serum, add undiluted positive control serum, 100μL/well. Others are sample wells, add the diluted sample, 100μL each.

2) Mix gently, cover and incubate at 37℃ for 30 min.

3) Remove adhesive foil. Pour the liquid out of the wells, add the diluted Washing buffer into each well fully, be static for 10s,pour out. Repeat 3 times, at last time pat to dry on absorbent paper.

4) Add 100μL enzyme conjugate into each well.

5) Cover plate with new adhesive foil. Incubate at 37 ℃ for 30 min.

6) Repeat step 3(washing).

7) Add substrate 100ul into each well, mix properly, incubate for 10 min at 37 ℃ in the dark with new adhesive foil.

8) Add stop solution 50μL into each well, mix gently and determine the result.

9) Measure the OD value of each well with a photometer at dual-wave length 450nm/630nm.

8. Results

For the assay to be valid, the positive control wells’ average OD value must be greater than or equal to 0.6, and the negative control wells’ average OD value is less than 0.1. Otherwise the test is invalid, need test again.

The result is judged by S/P value,

S/P=(Sample OD450/630- NCx(—))/( PCx(—)- NCx(—)), NCx(—) means Negative control’s average OD450/630 value(calculate as 0.05 when the value is less than 0.05), PCx(—) means Positive control’s average OD450/630 value

If S/P≥0.2, it is positive; less than 0.2, it is negative.

9. Precautions and warnings for users

1) This kit is for research use only.

2) Please read the manual carefully before use.

3) The experimental garbage was sterilized by high pressure steam at 121℃ for 30 minutes, or treated with 5.0g/L sodium hypochlorite disinfectant for 30 minutes and then discarded.

4) Microplates removed from refrigeration should be equilibrated with moisture, dried at room temperature, and ready to be opened. Return unused MicroWell plates to dry foil bags and seal at 4 °C. Unused liquid reagents should be covered and stored together with other components at 2-8°C away from light.

5) Samples and reagents should be added using a micropipette and frequently demonstrated for accuracy.

6) When adding washing buffer, it should be filled but not overflowed to avoid free enzyme or cross-contamination between wells.

7) The stop solution is corrosive, immediately rinse with plenty of water when it comes into contact with skin or clothes.

Specifications: 96 wells×2.

Expiry date: 12 months.

Storage: Store at 2~8℃, in the dark.

Production Date: On outer-packing of the test kit.

Q1: How long will it take for you to ship?
A1: Usually ships within 7 working days.

Q2: Do you support OEM/ODM?
A2: can be supported. We can customize according to your specific needs and specific quantities.

Q3: How is your factory doing in terms of quality control?
A3: We have ISO9001 certification and ISO13485 certification, so far all the production process, we have standard rules, we comply with the relevant behavior and laws of the government.

Q4: Is after-sales service guaranteed?
A4: We provide professional online technical after-sales service. If the product fails during the experiment, we can provide
one-to-one guidance through telephone, video and other forms.

Q5: What is your minimum order quantity?
A5: 1Kit.

Q6: What is the shipping method?
A6: Usually by air, we will choose the best shipping method for you, and we can also ship according to the shipping method you require.