Intended use of the testkit
This diagnostic test system for BLV infected cattle is intended to identify BLV p24 gp51 antibodies in individual or pooles serum samples. In contrast to test systems which make use of polyclonal antibodies or only one gp51 monoclonal antibody, this two monoclonal antibodies mediated ELISA gives a minimum of non-specific reactions.
Principle of the testkit
The principle of the test is based on the reaction of anti-BLV-p24 and anti-BLV-gp51 antibodies in test samples with BLV antigen which have been coated to a 96 well microtiter plate.
After washing, test serum samples are added to the wells and if present anti-BLV-p24 and anti-BLV-gp51 antibodies from the samples will bind to the antigen. The complex is detected by a horseradish peroxidase (HRPO) conjugated monoclobal antibody directed against bovine IgG. Color reaction in the wells is directly related to the presence of BLV-p24 and BLV-gp51 antibodies in the sample.
Interpretation of test results
In case of serum samples the following calculation should be used:
A sample is scored BLV negative if the OD value is below or equal to the average OD value of the negative control plus 0,250 OD units.
Negative: OD value samples < OD value negative control plus 0,250 OD units
In case of milk samples the following calculation should be used:
A sample is scored BLV negative if the OD value is below or equal to the average OD valua of the negative control plus 0.150 OD units.
Negative: OD samples < OD negative control plus 0.150 OD units.
If a sample does not meet these criteria for being negative, the following protocol is advised:
Serum samples: Confirmation of positive results can be achieved by testing individual serum samples in the EVL Bovine Leukaemia Serum complex-trapping-blocking ELISA test or PCR.