Porcine Toxoplasmosis IgG Antibody ELISA test kit

Porcine Toxoplasmosis IgG Antibody ELISA test kit

 

Porcine Toxoplasmosis IgG Antibody ELISA test kit
Catalog No. LSY-30001
1. Usage
Porcine Toxoplasmosis IgG antibody test kit is used for the detection of the Toxoplasmosis antibodies in porcine serum.
 
2. Principle
The test kit is enzyme immunoassay for the detection toxoplasmosis antibodies in porcine serum. Toxoplasmosis antigens are coated on 96-well plate, add sample into wells and then antibodies bound specifically by antigens, the unbound antibodies and other components will be removed by washing. Next add enzyme conjugation to specifically bind with the compound of antibodies and antigens on the microplate. The unbound enzyme conjugation will be removed by washing.Add the TMB substrate in the well, react to a blue product. At last, end the reaction by adding stop solution.
 
3. Components
1 Antigen coated microplate....................................... 2 pieces (96 wells each)
2 TOX-IgGNegative control serum (green lid)................................ 1.5 mL/tube
TOX-IgGPositive control serum (red lid)..................................... 1.5 mL/tube
3 Enzyme conjugate (yellow lid)..................................... 1 bottle (22 mL/bottle)
4 20×concentrated washing solution (white lid)............... 1 bottle (50 mL/bottle)
5 Substrate A (purple lid)/B (orange lid).................. 1 bottle each (12 mL/bottle)
6 Stop solution (blue lid)............................................... 1 bottle (12 mL/bottle)
7 Sample diluent solution (transparent lid)...................... 1 bottle (50 mL/bottle)
8 Adhesive Foil................................................................................. 6 pieces
9 Instruction....................................................................................... 1 piece
(Note: Never mix up reagents from test kits in different batch number)
 
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. 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. For single-wave length test, set one blank control well, add nothing; for double-wave length test, do not set blank control well. 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 Sample diluent solution, then add 1 μL serum sample separately (there will be color change after adding sample).
2          Mix gently, incubate at 37??? for 30 min.
3          Remove adhesive foil. Pour the liquid out of the wells, add Washing solution(dillute the Wash Concentrate at 20 times with deionized water) into each well fully, stand for 1 min. Repeat 5 times, at last time pat to dry on absorbent paper.
4          Add 100 μL enzyme comjugate into each well (except the blank well).
5          Cover plate with new adhesive foil. Incubate at 37 ??? for 30 min.
6          Repeat step 3(washing).
7          Add substrate A one drop (50 μL) and substrate B one drop (50 μL) into each well, mix properly, incubate for 15 min at 37 ??? in the dark with new adhesive foil.
8          Add stop solution one drop (50 μL) into each well, mix gently and determine the result within 5-30 min.
9          For single-wavelength test, measure the A value with a photometer at 450 nm, set zero for the blank well, and read A value of each well; For double-wavelength test, measure the A value with a photometer at 450 nm (Reference-wavelength: 630 nm), read A value of each well.
(Note: for single-wavelength test, if haven’t set zero at blank control well before reading, then the result should minus A value of blank control well)
 
7. Results
Cut-Off Value (C.O.) = 0.35
Generally speaking, the TOX-IgG Positive control OD mean should be greater than or equal to 0.70, the TOX-IgG Negative control OD mean should be less than 0.10, otherwise the test is invalid, need to retest.
If the sample OD value 0.35, the sample is classified as POSITIVE for TOX-IgG antibodies;
If If the sample OD value ???0.3, the sample is classified as NEGATIVE for TOX-IgG antibodies.
If between 0.3 and 0.35, it is skeptical.
 
8. 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.
 
9. Product performance
1. Specificity: to test 30 negative control serums, no false positive.
2. Sensitivity: to test 30 positive control serums, no negative.
3. Precision: CV???%???no bigger than 15%???n=10???
4. Stability: Store at 2??????8??? for 6 months or store at 37??? for 3 days, the result can reach the above 3 standards.
 
10. Precautions and warnings for users
1. This test kit is suitable for in vitro diagnostics.
2. Wear glove and working clothes when operate, treat the test kit as containing infectious material.
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. If the 20×concentrated washing buffer appears crystal, it is normal, put at 37??? until been dissolved.
6. Should use Micropipettor to add sample and reagents, and often proof its accuracy.
7. When adding washing buffer, should be full but no overflow, avoid appearing free enzyme at mouth of well or cross pollution between wells.
8. Stop solution is corrosive, use large amount of water to wash immediately when touch the skin or clothes.
 
Specifications: 96 wells × 2.
Expiry date: 6 months.
Storage:  at 2-8 ???, don’t expose in strong light, not freeze.
Production Date: On outer-packing of the test kit.
 
 
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@sybt.com  
www.lsybt.com
 

Product Origin: China
Model Number: LSY-30001
Brand Name: Green Spring
   
   

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Company Name: Shenzhen Lvshiyuan Biotechnology Co.,Ltd
Contact Person: Bella Zou
Address:
D building, National Biological Industrial Park of Marinelife, No.2 Binhai Road, Dapeng,  Shenzhen, China
Zip: 518120
Telephone:
+86-755-28438788
Fax: +86-755-28938800
E-mail: info@lsybt.com
Website: http://lsybtbella.tradeee.com

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