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LSY-30010 Livestock elisa Diagnostic kits PRRS Antibody ELISA assay kit

Categories Animal Disease ELISA Diagnostic kit
Place of Origin: China
Brand Name: Green Spring
Model Number: LSY-30010
Certification: ISO9001:2008 Certificate
Price: FOBShenzhen$180/kit
Packaging Details: By foam box with ice to maintain cool storage
Delivery Time: In 3 days after payment
Payment Terms: T/T or Western Union
Supply Ability: 500 kits/day
Properties: Diagnosis & Injection
Specifications: 96*2 wells/kit
MOQ: 1 kit
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    LSY-30010 Livestock elisa Diagnostic kits PRRS Antibody ELISA assay kit

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

    Catalog No. LSY-30010

    1. Brief

    The GreenSpring® Porcine reprodutive 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 reprodutive and respiratory syndrome virus in the pig farms; and investigation epidemiology of the PRRSV.


    2. Principle

    The GreenSpring® PRRSV Ab ELISA test kit is made from the antigen coated microtiter plate, goat-anti-pig IgG-HRP and other reagents. It applies the indirect ELISA principle to test the antibodies against PRRSV in porcine serum. In the test, the coated antigen combine with PRRSV-IgG in serum, then add IgG-HRP to specifically bind with complex of antibody-antigens 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 PRRSV-IgG.


    3. Components

    1

    PRRSV antigen coated microplate

    96T X 2

    2

    Enzyme conjugate

    22ml

    yellow lid

    3

    Sample diluent solution

    50ml

    transparent lid

    4

    PRRSV-IgG Negative control serum

    1.5ml

    green lid

    5

    PRRSV-IgG Positive control serum

    1.5ml

    red lid

    6

    Substrate

    12ml X2

    orange lid

    7

    Stop solution

    12ml

    blue lid

    8

    20×concentrated washing buffer

    50ml

    white lid

    9

    Adhesive Foil

    6 pieces


    10

    Instruction

    1 piece



    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.

    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 solution 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 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. (Can do for both single-well and double-well test).

    2 Mix gently for 10s, 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 comjugate 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 with new adhesive foil.

    8 Add 50 μL stop solution into each well, mix gently for 10s and determine the result.

    9 Measure the OD value with a photometer at 450 nm/630nm.


    8. ELISA analysis

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

    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. Limit of test method

    PRRSV-IgG can be used as the evaluation of PRRSV vaccine effect and PRRSV virus infection serological diagnostic indicators, but can not distinguish the two, if want to distinguish antibody caused by the vaccine or wild virus infection, it shout be combined with clinical data, immunization programs etc.


    11. 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 standards.


    12. Precautions and warnings for users

    1. This test kit is suitable for in vitro diagnostics.

    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.

    Specification

    Packing: 96 wells×2.

    Expiry date: 12 months.

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


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