Swinecheck APP 5a-5b
Immunoenzymatic assay for the detection of antibodies against Actinobacillus pleuropneumoniae serotypes 5a, 5b in porcine serum.
Pleuropneumonia caused by Actinobacillus pleuropneumoniae serotypes 5a, 5b is an important porcine disease that can result in considerable economic losses. Herds are often chronically infected without clinical symptoms. However, following a stressful situation, several clinical cases with mortality can occur. Since horizontal transmission through airways is quite important and infection is usually inapparent, serology becomes an indispensable tool for its detection.
Porcine serum samples and controls are diluted and incubated in wells coated with antigens (Ags) of Actinobacillus pleuropneumoniae serotypes 5a, 5b (APP 5a, 5b). The antibodies (Abs) specific to APP 5a, 5b present in positive serum samples will bind to the Ags in the wells. After several washes to eliminate unbound substances, a conjugate (an Ab coupled to an enzyme) targeted at the porcine Abs is added. After incubation, the excess of this conjugate is eliminated by a second wash and its attachment is revealed with a chromogenous substrate. Following this incubation, the enzyme, if present, reacts with the substrate and a green color develops. The reaction is then stopped and the optical densities are read. The intensity of the color allows the determination of the type of sample tested. A negative sample will show a weak reaction (pale green) whereas a strong positive will show a strong reaction (dark green). All shades of green between dark and pale represent various degrees of positivity.
Materials required but not provided:
- Purified water
- Adjustable single- and multi-channel micropipettes
- Single-use micropipette tips
- ELISA microplate washer
- Test tubes for sample dilution
- ELISA 96-well microplate reader equipped with 405 nm filter
- Containers for dilution of other solutions
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Reagents provided with the kit:
- 2 x 12 strips of 8 wells coated with APP 5a-5b antigens
- 1 x 2,5 ml Ready-to-use positive control
- 1 x 2,5 ml Ready-to-use negative control
- 1 x 50 μL Concentrated conjugate
- 2 X 100 mL Concentrated wash solution (10X)
- 1 x 24 ml Ready-to-use substrate
- 1 x 24 ml Ready-to-use stop solution
