Intended use of the test kit
This diagnostic test for leptospirosis is intended to identify antibodies against sugar antigens of Leptospira in serum and milk samples. In contrast to test systems which make use of un-purified non-specific Leptospira hardjo antigens, this test uses a monoclonal antibody which catches a specific Leptospira hardjo sugar antigen. This monoclonal based ELISA has very high specificity and sensitivity.
Principle of the test kit
An antigen solution antibody mixture is coated to the wells of the micro titer plate after stabilization and drying plates are vacuum sealed. Diluted milk or serum samples are added to the coated wells.
The bovine serum sample is added (diluted 1:100) to the wells of the coated plate.
The bovine milk sample is added diluted 1:3 to the wells of the coated plate.
The serum sample also can be titrated using a 3-step dilution, starting with a dilution 1:50 (> 1:150 > 1:450 > 1:1350).
The milk samples also can be titrated using a 3-step dilution, starting with a dilution 1:3 (> 1:9 > 1:27 > 1:81).
After incubation and appropriate washing, a monoclonal anti-bovine conjugate is added and the plates are again incubated.
After washing, substrate is added. Within several minutes the color reaction is stopped and the plates are immediately read at 450 nm.
Colour reaction in the wells is directly related to the concentration of the Leptospira antibody in the serum or milk sample.