Porcine Pseudorabies gB Antibody ELISA Test Kit 192 Wells/Kit GMP

1. Usage

The GreenSpring® Porcine Pseudorabies (PRV) Antibody ELISA Test Kit is used for detection of porcine Pseudorabies gB antibody in porcine serum; assessment of immunity conditions against porcine Pseudorabies in the pig farm and investigation of the epidemiology of the porcine Pseudorabies. While the result can not distinguish immune antibody and infectious antibody, to confirm that is antibody by vaccine or wild virus infection needs to combine clinical data, immune status, and other diagnostic methods.

2. Principle

The GreenSpring® Porcine Pseudorabies (PRV) gB antibody ELISA test kit is made from the antigen coated microtiter plate(coated with PRV antigen) and other reagents. It applies the Solid-phase ELISA principle to PRV-Ab in serum, then add enzyme conjugate to specifically bind with complex of coated antigen+PRV-Ab+enzyme labeled anti-pig-IgG antibody on the microplate. With the TMB substrate, it will generate an amount of color. The depth of color is relative with the content of the PRV-Ab, when the value of color is greater than the cut-off value, the pigs are vaccinated well or natural infected exist.

3. The 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) Microplate Washer.
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.
3) The EDTA, heparin sodiun and other anticoagulants will not affect the results.

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 at 40 times.(There will be color change after adding solution), 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.

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(both single- well and double-well test is OK).
2) Mix gently, cover andincubate 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, directly pour out, no need to be static. 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 ℃ for30 min.
6) Repeat step 3(washing).
7) Add substrate 100ul into each well, mix properly,incubate for 10 min at37℃ 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, 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. Interpretation of the result

1. Severe hemolysis, fiber protein in the serum separation is not sufficient, containing erythrocytes, a precipitate, a sample with bacteria may lead to false positive.
2. Negative results may occur on individual pigs after vaccines due to individual differences or immune duration.
3. Positive results for serological diagnosis and epidemiological investigation of swine to be combined with other methods and clinical data.

10. Product performance

1. Specificity: use this kit to detect reference serum, the compliance rate reach 100%.
2. Sensitivity: can reach max 1:5120.
3. Precision: CV(%)no bigger than 8%.
4. Stability: Store at 2℃～8℃for 12 months or store at 37℃for 3 days, the result can reach the above 3 standard.

11. Precautions and warnings for users

1. This test kit is for research use only.n.
2. Do not use reagents expired, do not mix reagents from different lots.
3. Read the Manual carefully before use.
4. 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.
5. 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.
6. If the 20×concentrated washing buffer appears crystal, it is normal, put at 37℃until been dissolved.
7. Should use Micropipettor to add sample and reagents, and often proof its accuracy.
8. When adding washing buffer, should be full but no overflow, avoid appearing free enzyme at mouth of well or cross pollution between wells.
9. Stop solution is corrosive, use large amount of water to wash immediately when touch the 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.

Epidemiology of Porcine Pseudorabies Virus

Pseudorabies virus is widely distributed in the world. Pseudorabies naturally occurs in pigs, cattle, sheep, dogs and cats. In addition, a variety of wild animals and carnivores are also susceptible. Mink and ferret can also cause pseudorabies outbreaks by feeding pig offal containing pseudorabies virus. Among experimental animals, rabbits are the most sensitive, and mice, rats and guinea pigs can also be infected. There are few reports on human infection with pseudorabies virus, and none of them is based on virus isolation.
Pigs are the storage host of pseudorabies virus, and diseased pigs, infected pigs and infected mice are important sources of infection of the disease. Many scholars believe that the infection of other animals is related to the contact with pigs and mice.
In pig farms, pseudorabies virus is mainly transmitted to healthy pigs through the elimination of infected pigs. In addition, the staff and equipment contaminated by pseudorabies virus play an important role in the transmission. Air transmission is the main way for pseudorabies virus to spread, but it is not clear how far it can spread. It is also found that cattle grazing around pig farms where pseudorabies occurs nearby can also get sick. In this case, air transmission is the only possible way. In pigs, the virus is mainly transmitted through nasal secretions. In addition, milk and semen are also possible ways of transmission.
After infected with pseudorabies virus, all animals except pigs died.
Pseudorabies occurs seasonally, mostly in cold seasons, but also in other seasons.

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: It can be shipped by express (FEDEX, UPS, DHL, EMS, etc.) or by air and ground.Please confirm with us before placing an order.

Shenzhen Wensidun Technology Co., Ltd. was established in 2018 and is located in Longgang District, Shenzhen, Guangdong Province, China. We have in-depth cooperation with Green Spring, which has 20 years of production and R&D experience.

We have more than 2000 square meters of GMP clean production workshop and more than 1000 square meters of R&D center in Shenzhen. Products cover over 100 varieties of food safety test kits, animal diseases diagnostic kits, in vitro diagnostic reagents, genetic antibody antigen preparation and form a complete set of testing instrument.Currently,our food safety test kit and animal disease diagnostic kit are widely used in many governments,enterprises and research institutes.Our products are sold to more than 40 countries and areas.

We have ISO9001, ISO13485 quality management system certification, GMP certificate. SARS-CoV-2 Antigen Rapid Test Kit has been awarded CE certification, German BfArM approved self-test and professional test, PEI certification, EU general and mutual recognition list, and multinational white list. Ranked No. 1 in German authoritative medical magazine Daz and No. 1 in Hong Kong sales.

Enterprise mission: Caring for health and benefiting the society. Focus on products and lead the industry.

Core values: Inheriting innovation, improving quality, honesty and law-abiding, and win-win cooperation.

Our vision is to become a leading company in the biodiagnostic reagent industry, providing high-quality, convenient and reliable services to the world.