Enteritis Ag (Rota, Corona, K99)
Immunoenzymatic assay intended for the detection of rotavirus, coronavirus and Escherichia coli (E. coli) K99 antigens in bovine feces.
Diarrhea problems in the calf are often caused by rotavirus, coronavirus, or E. coli K99. These problems can lead to important economical losses since affected animals generally show reduced weight gain, sometimes resulting in death. Quick identification of the pathogen involved helps in disease control. It is recommended to test the feces of at least 3 calves per infected herd throughout symptom evolution.
Feces samples are diluted and incubated in wells coated with a mixture of 3 antibodies (Abs) specific to rotavirus, coronavirus, and E. coli K99. If one of these pathogens is in the feces samples, it will bind to its specific Ab already in the wells. After several washes to eliminate unbound substances, a conjugate (an Ab coupled to an enzyme) targeted at either rotavirus, coronavirus, or E. coli K99 is added. After incubation, excess of this conjugate is eliminated by a second wash and its attachment to the specific pathogen is revealed with a chromogenous substrate. Following this incubation, the enzyme, if present, reacts with the substrate and a blue color develops. The intensity of the color allows the determination of the type of sample tested. A negative sample will show a weak reaction (colorless, pale blue) whereas a positive sample will show a reaction stronger than the one for the negative control (darker blue than negative control).
Materials required but not provided:
- Purified water
- Test tubes for sample dilution
- Containers for dilution of other solutions
.
Reagents provided with the kit:
- 1 x 12 Strips of 8 wells coated with a mix of 3 Abs specific to rotavirus, coronavirus, and E. coli K99
- 1 x 2 ml Ready-to-use positive control (dark yellow)
- 1 x 2 ml Ready-to-use negative control (pale yellow)
- 1 x 200 ml Ready-to-use dilution buffer
- 1 x 20 ml Concentrated wash solution (50X)
- 1 x 8,5 ml Ready-to-use anti-rotavirus conjugate (red)
- 1 x 8,5 ml Ready-to-use anti-coronavirus conjugate (blue)
- 1 x 8,5 ml Ready-to-use anti-E. coli K99 conjugate (green)
- 1 x 14 ml Substrate A
- 1 x 4 ml Substrate B
- 1 x 3 ml pipette
- 30 x 1 ml pipettes
