Swinecheck HPS (Haemophilus parasuis)
Immunoenzymatic assay for the detection of antibodies against Haemophilus parasuis (HPS) in porcine serum regardless of the serotype.
HPS is a commensal organism of the upper respiratory tract of pigs. Fifteen different serotypes have been identified so far. Serotypes 1, 5, 10, 12, 13 and 14 are the most virulent. Under appropriate conditions virulent strains can cause a severe systemic disease known as Glässer’s disease. Clinical signs and lesions vary depending on the immune status of the animals and the virulence of the strain. Systemic infection of naive animals with virulent HPS strains is characterized by high fever, lethargy, swollen joints, coughing, dyspnoea, and central nervous system signs, such as trembling and paddling. Glässer’s disease may be controlled with antimicrobials and prevented by vaccination. Diagnosis of HPS infections is based on clinical signs, lesions at necropsy, and detection of the organism by isolation or PCR. Detection of HPS circulating antibodies may be useful to diagnose HPS infection in animals which have demonstrated suspect clinical signs or to evaluate levels of maternally derived antibodies prior to vaccination or antibody response after vaccination.
The diluted porcine serum samples and the controls are incubated in wells coated with HPS antigens (Ag) (odd colums), and in wells without Ag (even columns). The antibodies (Ab) specific to HPS present in positive serum samples bind to the Ag in the wells. After several washes to eliminate unbound substances, a conjugate (an Ab coupled to an enzyme) targeted at porcine Ab 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 status 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 provided
- 12 Strips of 8 wells coated with HPS Ag
- 12 Strips of 8 wells without HPS Ag used as control (blue sticker)
- 1 x 2 ml Ready-to-use positive control
- 1 x 2 ml Ready-to-use negative control
- 1 x 50 μL Concentrated conjugate
- 2 x 100 ml Concentrated wash solution 10X
- 1 x 30 ml Ready-to-use substrate
- 1 x 30 ml Ready-to-use stop solution
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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
