Swinecheck Influenza H1N1
Immunoenzymatic assay for the detection of swine influenza virus, subtype H1N1, antibodies in porcine serum.
The influenza virus is responsible for swine flu, one of the most common respiratory diseases throughout the world. Swine flu is contagious and mainly characterized by coughing, 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 will optimize vaccine selection.
Porcine serum samples are diluted and incubated in wells coated with H1N1 antigens (Ag). The antibodies (Ab) specific to H1N1 in positive serum samples will bind to the Ag in the wells. After several washes to eliminate unbound substances, a monoclonal antibody (Mab), also specific to the H1N1 Ag, is
added. The Mab binds to the Ag that are not yet bound by the Ab found in the previously added serum samples. After several washes to eliminate unbound substances, a conjugate (an Ab coupled to an enzyme) targeted at the Mab is added. After incubation, the excess of this conjugate is eliminated by another wash and its attachment is revealed with a chromogenous substrate. Following this incubation, the enzyme, if present, reacts with the substrate and a blue color develops. The reaction is then stopped (the color changes from blue to yellow) and the optical densities are read. The intensity of the color indicates the status of the sample tested. A strong positive is indicated by a weak reaction (pale yellow). Conversely, a strong reaction (dark yellow) indicates negative samples. Each shade of yellow represents varying degrees of positivity.
Materials Provided:
- 2 x 12 strips of 8 wells coated with H1N1 Ag
- 1 x 1,5 ml Ready-to-use Positive Control
- 1 x 1,5 ml Ready-to-use Negative Control
- 1 x 50 ul Concentrated Monoclonal Antibody (400X)
- 1 x 200 ul Concentrated Conjugate (75X)
- 1 x 15 ml Ready-to-use Conjugate Dilution Buffer
- 1 x 100 ml Concentrated Wash Solution (25X)*
- 1 x 30 ml Ready-to-use Substrate
- 1 x 30 ml Ready-to-use Stop Solution
.
Materials Required but not Provided:
- Purified water
- Adjustable single- and multi-channel micropipettes
- Single-use micropipette tips
- Test tubes for sample dilution
- ELISA 96-well microplate reader equipped with 450 nm filter
- Containers for the preparation of solutions
