Intended use of the test kit
This diagnostic test is intended for the detection of rabies virus nucleoprotein in inactivated antigen samples. This standardized ELISA is based on monoclonal antibodies, with high sensitivity and specificity for the nucleoprotein of rabies virus. This testkit was used in the collaborative study organized by the National Institute of Health and Environmental Protection (RIVM, The Netherlands).
Principle of the test kit
The test is based on the reaction of rabies nucleoproteins with biotinylated monoclonal antibodies. To this end rabies nucleoproteins have been coated to a 96-well microtiter strip plate.
The antigen samples are added to the wells of a pre-incubation plate. Immediately the same volume of a biotinylated monoclonal antibody solution (MAb conjugate) is added.
After pre-incubation the antigen/conjugate mixture is transferred to the coated microtiter plate to bind un-complexed MAb conjugate. After washing, the bound MAb conjugate is detected by Avidin HRPO conjugate.
The antigen sample is added (diluted 1:8) to the wells of the coated plate.
The antigen sample also can be titrated using a 2-step dilution, starting with 1:2 (> 1:4 > 1:8 > 1:16 > 1:32 > 1:64 > 1:128 > 1:256).
After developing with substrate, the color reaction in the wells is reversed related to the concentration of rabies virus nucleoprotein in the sample.