AI Article Synopsis
- The indirect ELISA was created to detect antibodies against BHV-5 in sheep sera, using a specific antigen preparation that was purified effectively through a sucrose gradient.
- Optimal testing conditions included coating the solid phase at 4°C overnight with virus material, and background interference was minimized using a blocking agent.
- Out of 490 sheep sera samples tested, a threshold was established for distinguishing positive from negative results, highlighting the rising importance of monitoring BHV infections due to their economic impact on ruminants.
Article Abstract
The indirect enzyme-linked immunosorbent assay (ELISA) was first developed for the detection of antibodies against BHV-5 in sheep sera. Investigations of various antigen (Ag) preparations revealed that BHV material which had been purified across a sucrose gradient using sonification followed by ultracentrifugation gave satisfactory results during application in ELISA. Optimal coating of the solid phase was achieved with Ag adsorption at 4 degrees C overnight with 5 micrograms of virus material/ml in a carbonate-bicarbonate-buffer, pH 9.7. The background reactions were significantly reduced by blocking the free bonding points with 1% gelafusal after Ag adsorption. As the extinction values of all the sera investigated which a dilution of 1:50 or more below 0.2, the difference threshold between positive and negative sera was set at 0.2. A total of 490 sera from various stocks of sheep were investigated with regard to the occurrence of antibodies against BHV-5 in ELISA. The growing rate of infection with herpesviruses among ruminants and the subsequent economic loss have resulted in the increased significance in the last few years of bovine herpes-virus (BHV) disease.
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