Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Antibody ELISA kit

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Antibody ELISA kit

Catalog No. LSY-30010

1. Brief

The GreenSpring^® Porcine reproductive and respiratory syndrome virus antibody test kit (PRRSV Ab) is used for the detection of PRRSV antibodies in swine serum; assessment of immunity conditions against porcine reproductive and respiratory syndrome virus in the pig farms; and investigation epidemiology of the PRRSV.

2. Principle

The GreenSpring® PRRSV antibody ELISA test kit uses the principle of solid-phase enzyme-linked immunosorbent assay (ELISA), and consists of a microwell reaction plate coated with high-purity PRRSV antigen, anti-pig IgG labeled with horseradish peroxidase, and other reagents. The reaction mechanism is that the coated antigen is combined with the PRRSV-Ab in the sample, and then combined with the enzyme-labeled anti-pig IgG antibody to form a "coated antigen+PRRSV-Ab+enzyme-labeled anti-pig IgG antibody" complex. Add adding Substrate, Catalytic reaction color development. The color depth is directly proportional to the amount of PRRSV-Ab. When the color reaction of the sample is detected, the result obtained by the microplate reader exceeds the set threshold, and the result is positive, which indicates that the immune has produced antibodies or there is a natural infection.

3. Components

4. Material Required not Provided

1. Microplate Reader (Dual-wave length: 450/630 nm).
2. Microplate Washer.
3. Micropipettes, adjustable (Single-wave length 1-100ul,0.5-10ul,multi-wave length 30-300ul).
4. Constant temperature box or water bath box.
5. Oscillator.
6. Disposable tips (10ul, 200ul)
7. 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.

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

3) Washing solution preparation: Dilute the 10×concentrated washing buffer with deionized water at 10 times. (eg.10ml 10×concentrated washing buffer + 90ml deionized water ), if there is crystallization in the 10×concentrated washing buffer, it is normal, dissolve it at 37℃.

7. Procedure

1. Adding sample: Take out the required coated plates according to sample quantity (Can be detached) and record the sample position on a worksheet. Set 2 wells for negative control serum and 2 wells for positive control serum, add undiluted negative and positive control serum to its well accordingly, 100 μL/well. Others are wells for samples, add 100μL/well of the diluted Sample.
2. Cover plate with adhesive foil, incubate at 37℃ for 30 min.
3. Remove adhesive foil. Pour the liquid out of the wells, add Washing solution into each well fully, be static for about 10s, pour out directly. Repeat 3 times, at last time pat to dry on absorbent paper.
4. Add 100 μL 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 100ul substrate into each well, mix properly, incubate for 10 min at 37 ℃ in the dark.
8. Add 50μL stop solution into each well, mix gently for 10s and determine the result.
9. Measure the OD value with a photometer at 450nm/630nm.

8. ELISA analysis

Measure the OD Value of each well, for the assay to be valid the following specifications must be met. The average OD value of PRRSV-Positive control must be equal to or greater than 0.60, the average OD value of PRRSV-Negative control must be less than 0.15.

The result is judged by S/P value,

S/P=(Sample OD_450/630- NCx(—))/( PCx(—)- NCx(—)), NCx(—) means Negative control’s average OD_450/630 value, PCx(—) means Positive control’s average OD_450/630 value

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

9. Precautions and warnings for users

1. Read the Manual carefully before use.

2. Do not use reagents expired, do not mix reagents from different lots.

3. Experiment rubbish should be dealt with high pressure steam sterilization at 121 ℃ for 30 minutes, or treated with 5.0g/L sodium hypochlorite disinfectant for 30 minutes, then discard.

4. 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 4 ℃. Unused liquid reagent should cover caps, store at 2-8 ℃ in dark with other group components.

5. Should use Micropipettor to add sample and reagents, and often proof its accuracy.

6. When adding washing buffer, should be full but no overflow, avoid appearing free enzyme at mouth of well or cross pollution between wells.

7. Stop solution is corrosive, use large amount of water to wash immediately when touch the skin or clothes.

Specification

Packing: 96 wells×2.

Expiry date: 12 months.

Storage: Storing at 2~8 ℃in the dark, not freeze.

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