IMMUNOPEROXIDASE TEST FOR VIROLOGIC DIAGNOSTICS OF PORCINE CIRCOVIRUS INFECTION

摘要

An immunoperoxidase test for virologic diagnostics of porcine circovirus infection involves inoculation of sensitive culture cells onto 96-cell plastic microplate with flat bottom, dilution of test virus-containg material by ten-fold dilution technique in 96-cell plastic microplates with round bottom followed by transferring thereof into microplate cells with monolayer of cell culture, incubation in COconditions of temperature-regulated chamber for 7 days, removal of culture medium from microplate cells with washing monolayer of cell culture, addition of fixing buffer into all microplate cells, stepwise addition of working dilution of specific blood serum, working dilution of conjugate, substrate solution into all microplate cells, termination of reaction by addition of stop solution into each microplate cell and reaction recording by means of inverted light microscope wherein infected cells have the appearance of isolated cells or cell clumps with deep red colored cytoplasma and pale nucleus. The method is used for virologic diagnostics of porcine circovirus infection in domestic and wild pigs by investigation of biological material (blood, suspension of organs, biological fluids) or by determination of infectious activity titer of vaccine, reference strains or field isolates of II type circovirus.