This study produced the pseudorabies virus (PrV) high immune serum by multi-incubated the pseudorabies antibody negative healthy pig with strong pseudorabies vaccine strain S with different doses at certain intervals. And identified the high immune serum by neutralization test, ELISA and immuogold labeling test strip. The results approved that the PRV high immune serum produced by this method is of high specification, the titer is over 256 times, completely meet the needs of the diagnostic requirements.