Swinecheck Influenza H3N2
Immunoenzymatic assay for the detection of antibodies against swine influenza (H3N2) in porcine serum.
The influenza virus is responsible for swine flu, one of the most common diseases throughout the world. Swine flu is contagious and mainly characterized by cough, anorexia, lethargy, and nasal discharge. Other characteristics, although seldom observed, include abortion for sows and death for piglets. This virus has several subtypes, the most common being H1N1 and H3N2. The differentiation between the subtypes is important since it can help in selecting the vaccine.
Porcine serum samples are diluted and incubated in two wells. The first wells (odd columns) are coated with antigens (Ags) of the swine influenza virus (H3N2). The second wells (even columns) contain no Ags and serve as control to ensure that the positive reactions observed in the wells containing Ags are truly the result of an interaction between the porcine serum antibodies (Abs) and the H3N2 specific Ags. The Abs specific to H3N2 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 Provided:
- 1 x 12 Strips of 8 wells coated with H3N2 Ags
- 1 x 12 Strips of 8 wells without H3N2 Ags used as control (blue)
- 1 x 2.5 ml Ready-to-use positive control
- 1 x 2,5 ml Ready-to-use negative control
- 1 x Concentrated conjugate (Volume may vary)
- 1 x 125 ml Dilution buffer A
- 1 x 1,25 ml Dilution buffer B
- 1 x 80 ml Concentrated wash solution (25X)*
- 1 x 25 ml Ready-to-use substrate
- 1 x 25 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
