Swine Mycoplasma Pneumonia antibody ELISA kit Veterinary test

Swine Mycoplasma ELISA kit

Catalog No. LSY-30022

1. Brief

Swine mycoplasma is a smallest self-propagation prokaryote between viruses and bacteria. It is widely distributed in nature, can infect human and animal. Mycoplasma infection can cause multiple system disease in animal. By laboratory methods, early and timely detection of Mycoplasma have important value. The kit can be used to detect specific antibody of Mycoplasma in pig serum.

2. Components

1. Pre-coated Microplate.............................................……1 pieces (96 wells)
2. Enzyme Conjugate (No.1 solution)..........................................1 bottle 12ml
3. 10x Concentrated washing solution (No.2 solution)................1 bottle 100ml
4. Substrate (No.3 solution) ......................................................1 bottle 15ml
5. Color reagent (No.4 solution)………………………………………1 bottle 15ml
6. Sample diluent (No.5 solution)……………………………………1 bottle 100ml
7. Stop solution (No.6 solution) ………………………………………1 bottle 15ml
8. Positive control.....................................................................1 bottle 1ml
9. Negative control ...................................................................1 bottle 1ml
10. Adhesive film.............................................................................1 /2 piece
11. Sealed bag....................................................................................1 piece
12. Instruction......................................................................................1 piece

3. Materials required but not provided

ELISA Reader including 450nm and 630nm wavelength, 37℃ Incubator, measuring pipettes.

4. Sample requirement

1) To obtain best laboratory result, avoid using sample with NaN3,high cholesterol,bacteria and severe hemolytic.

2) Store serum sample at 2~8℃ for a short term(7 days); store at -20℃ or below -20℃ for a long time, avoid repeated freezing and thawing.

5. Procedure

1). Washing buffer preparation: dilute concentrated washing solution (No.2 solution) with distilled water or deionized water at 1:10(such as 100ml concentrated washing solution+900ml distilled water or deionized water), mix thoroughly to obtain the washing working solution.

2). Sample dilution: Dilute serum sample with sample diluent (No.5 solution) at 1:100 (such as 5ul serum sample + 495ul sample diluent), mix thoroughly . Negative control and positive control do not need dilute, add directly.

3). Adding sample: for each test, set 2 wells for negative control, add negative control 100ul /well, 1 well for positive control, add positive control 100ul /well, 1 blank control well, add 100ul sample diluent(No.5 solution). For other wells, add 100ul diluted serum sample. Incubate at 37 ℃ in dark for 30 min, pour the liquid out of the wells. Use the diluted washing solution to wash for 3 times, each time stand for 1 min, at last time pat to dry on absorbent paper.

4). Adding enzyme conjugate: Except the blank well, add 1 drop of enzyme conjugate (No.1 solution) to each well.Incubate at 37 ℃ in dark for 30 min, pour the liquid out of the wells. Wash as last step, pat to dry on absorbent paper.

5). Coloration: Add 1 drop of Substrate (No.3 solution) and Color reagent (No.4 solution) accordingly, mix evenly,Incubate at 37 ℃ in dark for 10 min.

6). Add 1 drop of Stop solution (No.6 solution) to stop reaction (blue will change into yellow after adding stop solution) and determine the result.

6. ELISA analysis

Set zero for the blank well, measure with microplate reader at 450nm (Reference-wavelength: 630 nm).

1. Valid determination:

OD Value of Negative control ≤0.15 (the test is invalid when >0.15)

OD Value of Positive control ≥0.30 (the test is invalid when <0.30)

1. Result judgment

When Average OD value of Negative control is less than 0.08, calculate as 0.08,

S/N= OD Value of Samples/ Average OD value of Negative control

S/N > 2.1, swine mycoplasma is positive;

S/N ≤ 2.1, swine mycoplasma is negative.

4) Limitation of the test method: This test kit is only applied to swine serum.

7. Note

1) Return to room temperature before use. Seal the unused micro-well strip and store at 2~8 ℃.

2) Each reagent must be shaken evenly before use.

3) Seal the plate with adhesive film when incubate, do not repeat use the adhesive film.

4) Never mix up reagents from test kits in different batch number. Tighten all caps after use, don’t mix caps.

5) The wasted kits and sample should be treated as infection agent. The stop solution is corrosive, be careful to use.

Specifications:96 wells

Expiry date:12 months.

Storage:Store at 2~8℃ in dark, no frozen.