Product For Pig

LSY-30041 Actinobacillus Pleuropneumoniae(APP) ApxIV antibody ELISA kit for Swine

Actinobacillus Pleuropneumoniae(APP) ApxIV antibody ELISA kit for Swine

Catalog No. LSY-30041

1. Usage andPrinciple

This kit use indirect enzyme-linked immunoassay (indirect ELISA) to detect specific Actinobacillus Pleuropneumoniae(APP) ApxIV antibody in swine serum or plasma. Applicable to the differential diagnosis of Actinobacillus Pleuropneumoniae infected pigs and inactivated vaccines or subunit vaccines Immune pigs. 

If there is Actinobacillus Pleuropneumoniae(APP) ApxIV antibody in sample, it will combine with coated Actinobacillus Pleuropneumoniae(APP) ApxIV antigen on plate and enzyme conjugate added on second step. The color reaction of anti-porcine enzyme conjugate on plate with substrate will reflect whether the sample is infected with Actinobacillus Pleuropneumoniae(APP) .

2. Reagents and contents

Code

Item

Spec.

Code

Item

Spec.

1

ApxIV-AgCoatedplates

96 wells*2

7

Stop solution

15 ml

2

EnzymeConjugate

22 ml

8

Positive control

1ml

3

10X Washing solution

100 ml

9

Negative control

1ml

4

Substrate A

15 ml

10

Adhesive Foil

6 pieces

5

SubstrateB

15 ml

11

Instruction sheet

1 piece

6

Sample dilution 

100 ml

12

Serum dilutionplate

2 piece

3.Materialrequired notprovided

1) Microplate Reader with 450nm and 630nm

2) 37 ℃ thermostatic device

3) Micropipettes, adjustable.

4. Samplerequirement

1) This kit only used to testing porcine serum or plasma.

2) To get the best test result, avoid using sample with severe hemolysis, precipitate, contaminated by bacteria or protein suspension.

3) The serum sample can store at 2-8 ℃for short term (in 3 days),  if for long term, it should be kept at -20℃or lower, avoid repeated freezing and thawing.

5.Test procedure

1) Washing Solution preparation: Dilute 10X Washing solution with distilled water or deionized water at 1:10 (for example: take 100ml 10X Washing solution, add 900ml distilled water or deionized water, mix),mix it evenly to get washing solution.  

2) Sample dilute and adding sample: Dilute serum sample with sample dilution at 1:40, add only diluted serum sample to sample wells, 100ul/well.If use Serum dilution plate to dilute sample, the mothed is as following: add 195ul Sample dilution into Sample dilution plate, then add 5ul serum, use Pipettes to blow and stir to mix it evenly, then take 100ul solution into the sample wells. (Note:the Serum dilution plate is disposable, do not repeat use.)  

3) Adding controls: Negative control and positive control do not need dilute, add it directly. Set 2 wells for blank control, only add 100ul sample dilution to each well; 2 wells for negative control, only add 100ul negative control to each well; 2 wells for Positive control, only add 100ul Positive control to each well. Cover plate with adhesive foil,incubate in dark at 37℃for 30 minutes.

4) Washing plate: discard the liquid of the well, fill full (add 250ul/well if using washing machine)with washing solution to each well, washing for 3 times, be static for 1 min for each time; at last time, flap to dry with the absorbent paper.

5) Adding Enzyme Conjugate: Except blank control, add Enzyme Conjugate to each well, 100ul/well, Cover plate with adhesive foil,incubate in dark at 37℃for 30 minutes. Discard liquid of the wells and wash 3 times as described in step 4).

6). Adding substrates: according to the quantity needed, take same volume of Substrate A and Substrate B, mix them evenly before use, then add into all well,100ul/well, Cover plate with adhesive foil,incubate in darkat 37℃ for 10 minutes.

7). Stop reaction: Add 50ul Stop solution into all wells to stop reaction. Set zero at blank control, read the OD value at ELISA reader 450nm (630nm as a reference).


6. Reference

Validation judge

Negative control OD value≤0.20 (If more than 0.20, the test is invalid)

Positive control OD value ≥ 0.50 (If less than 0.50, the test is invalid)


7. Resultsinterpretation

S/P= OD value of sample/ Average OD value of Positive control

S/P value≥0.2: Actinobacillus Pleuropneumoniae(APP) ApxIV antibiody Positive;

S/P value < 0.2: Actinobacillus Pleuropneumoniae(APP) ApxIV antibiody Negative.

Limitation: This kit is for screening only, Not as a basis for diagnosis.


8. Notes

1) 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 2~8 ℃.

2) Before adding reagent, gently shake dropping bottle and mix even the reagent.

3) When incubation, must cover the plate with adhesive foil, do not repeat use the adhesive foil.

4) Don't mix use components and instruction from different kits

5) The test kit and any samples should be regarded as the source of infection to properly handle. Stop solution is corrosive, be careful to use.

9. Storage and expire date

Store at 2~8℃ in dark, no frozen, expiry date: 12 months.


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Company: Shenzhen Lvshiyuan Biotechnology Co.,Ltd

WeChat/WhatsApp: +86-13427908554

Mobile: +86-18165709090 Skype: bellazou3

E-mail: info@lsybt.com, lsy@lsybt.com

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