AffiVET® Bovine Brucella Antibody C-ELISA Kit
Species: Ruminant
Disease or Pathogen: Brucellosis
Target: Antibody
Test Type: Elisa Kit
Principle of the Test
Brucellosis is a highly contagious bacterial disease, almost exclusively caused by Brucella spp. causing late term-abortion and infertility in cattle. The disease is also a serious zoonosis, causing undulant fever in humans.
The AffiGen Brucella Ab C-ELISA is a competitive Enzyme Linked Immunosorbent Assay(ELISA) for the qualitative detection of Brucella spp. antibody in serum or plasma. This is able to differentiate between vaccination antibodies(S19 vaccine) and Brucella infection in cattle.
The AffiGen Brucella Ab C-ELISA contains a microplate, which is pre-coated with non-infectious Brucella antigen on the well. For testing, AffiGen Brucella Ab C-ELISA plates are incubated with an equal mixture of sample and monoclonal antibody-Biotin for 30 minutes at the Room temperature. During first incubation, Brucella antibodies(if present in the test sample) and anti-Brucella LPS monoclonal antibody bind competitively to the antigens in the well. After incubation, Unbound materials are removed by aspiration and washing. Streptavidin-HRP conjugate added subsequently forms a complex with the monoclonal brucella antibody-Biotin cunjugates.
Unbound materials are removed by aspiration and washing again and substrate solution is added. The enzyme activity found in the well will thus be directly inversely proportional to the Brucella antibody concentration in samples. The reaction is stopped by addition of the stopping solution and colorimetric reading should be performed by using a spectrophotometer at 450 ㎚ with reference wavelength at 620 ㎚.
Materials provided
AffiGen Brucella Ab C-ELISA contains following items to perform the assay.
| Microplate coated with LPS of Brucella spp. | 1 plate (96wells/plate) configured in twelve 1x8 strips. |
| Negative control(NC) serum | Normal bovine serum and Proclin 300 (0.05%) |
| Strong Positive Control(SPC) serum | Strong Positive bovine serum against Brucella antibody, Proclin (0.05%) |
| Weak Positive Control(WPC) serum | Weak Positive bovine serum against Brucella antibody, Proclin (0.05%) |
| Sample diluents (5X concentrated) | Phosphate buffered saline, Proclin (0.01%) |
| Amplification Conjugate (3X concentrated, Freeze dried) | Anti-Brucella antibody Biotin conjugate |
| Washing solution (20X concentrated) | PBS-Tween 20, Proclin (0.05%) Note: If undissolved crystals are present, re-suspend the solution by placing the bottle at 37℃ for few minutes. |
| Enzyme conjugate (101X concentrated) |
Strepavidin-HRP counjugate, BSA, Proclin (0.05%) |
| Conjugate diluents | Phosphate buffered saline, Proclin (0.05%) |
| Substrate (Ready to use) | Tetramethyl-benzidine with citrate-phosphate buffer containing H2O2. |
| Stopping solution (Ready to use) | 1N sulfuric acid. Ready to use. |
| Adhesive plate sealer |