Ref. CDVB015
96 tests/kit
FASCIOLA HEPATICA
Bovine
General Information
Fascioliasis is a disease of livestock caused by the parasitic
trematode Fasciola hepatica. In cattle that harbor patent infections of
Fasciola, the recovery of eggs from their feces remains the best method of
diagnosis of liver fluke infection. However, symptoms due to acute
fascioliasis occur much earlier and the parasite eggs often are not found
even after repeated stool examinations.
The enzyme-linked immunosorbent assay (Elisa) has been adapted recently for the diagnosis of fascioliasis in order to avoid the time-consuming coprological methods. The excretory-secretory antigens have been most widely adopted for the immunodiagnosis of fascioliasis because they have both immunodiagnostic and immunoprophylactic potential.
The Cypress’ Fasciola hepatica Elisa kit allows the reliable detection of antibodies to the parasite in infected cattle.
Principle of the test
The Cypress Fasciola Hepatica test uses 96-well microtitration plates
sensitized by a specific Fasciola hepatica antigen.
The plates’ odd columns (1,3,...11) contain the specific antigen, whereas the even columns (2,4,.....12) are used to control the specificity of the test because they do not contain the antigen. We thus have a genuine negative control to differentiate the specific anti-Fasciola hepatica antibody from the non specific antibodies. Using such a control reduces the number of false positives considerably.
The test blood sera are diluted 1 : 100 in the buffer for dilution. The plate is incubated and washed, then the conjugate - a peroxidase labeled anti-Bovine IgG1 monoclonal antibody - is added to the wells. The plate is then incubated a second time at room temperature and washed again and the enzyme’s substrate (hydrogen peroxide) and the chromogen tetramethylbenzidine (TMB) are added. This chromogen has the advantages of being more sensitive than the other peroxidase chromogens and not being carcinogenic.
If specific Fasciola hepatica immunoglobulins are present in the test sera the conjugate remains bound to the microwell that contains the antigen and the enzyme catalyses the transformation of the colorless chromogen into a pigmented compound. The intensity of the resulting blue color is proportionate to the titer of specific antibody in the sample. The signal read off the negative control microwell is subtracted from that of the positive microwell sensitized by the antigen.
Comparison of the signals obtained for the unknown samples with those obtained for a reference serum, supplied with the kit, make it possible to rank the serums positivity on a five point scale.
Rev.09.95
