Medycyna Wet. 63 (6), 712-716, 2007
Larska M., Rola J.
Application of the indirect immunoperoxidase test to confirm the specificity EAV isolates
The purpose of the study was the elaboration and introduction of an effective method to confirm the identity of EAV isolates obtained in cell culture from the serum of persistently infected stallions. In view of the low cost, simplicity of preparation, and its high specificity, an indirect immunoperoxidase test based on rabbit polyclonal sera was introduced. After immunization of the rabbits with the EAV reference strain Bucyrus, purified by ultracentrifugation, a specific polyclonal serum in the titer of SN antibodies from 1:32 to 1:128 was obtained. Peroxidase-conjugated goat IgG antibodies against horse immunoglobulins were used as a secondary antibody and DAB as a substrate. After the optimalization of the test conditions, its sensitivity and specificity was defined. The sensitivity estimated in reference to the Bucyrus strain was equal to 1 TCID50. The specificity was defined at 100%. After the preliminary validation, the indirect immunoperoxidase test based on the obtained polyclonal sera was affirmed as a method to identify EAV isolates. All 68 EAV field isolates reacted positively in the immunoperoxidase test.
Keywords: equine arteritis virus, indirect immunoperoxidase test