Actinobacillus Pleuropneumoniae(APP) ApxIV antibody ELISA kit for Swine
Catalog No. LSY-30040
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) .
|1||ApxIV-Ag Coated plates||96 wells*2||7||Stop solution||15 ml|
|2||Enzyme Conjugate||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||Substrate B||15 ml||11||Instruction sheet||1 piece|
|6||Sample dilution||100 ml||12||Serum dilution plate||2 piece|
3. Material required not provided
1) Microplate Reader with 450nm and 630nm
2) 37 ℃ thermostatic device
3) Micropipettes, adjustable.
4. Sample requirement
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
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
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 dark at 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).
Negative control OD value ≤ 0.20 (If more than 0.20, the test is
Positive control OD value ≥ 0.50 (If less than 0.50, the test is
7. Results interpretation
S/P= OD value of sample/ Average OD value of Positive control
S/P value ≥ 0.2: Actinobacillus Pleuropneumoniae(APP) ApxIV
S/P value < 0.2: Actinobacillus Pleuropneumoniae(APP) ApxIV
Limitation: This kit is for screening only, Not as a basis for diagnosis.
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
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.
Shenzhen Lvshiyuan Biotechnology Co., Ltd
D Building, National Biological Industrial Park of Marinelife, No.2
Binhai Road, Dapeng, Shenzhen, 518120 China