Experimental data show the usefulness of the Leghorn chicken as a producer of the enterovirus neutralizing antibodies (IgY). The resulting serum is not inferior to the specific activity of the commercial rabbit enterovirus diagnostic sera (EDS) in the neutralization reaction. The IgY have lower backgrounds than mammalian IgG and do not cause toxic effect to cell culture. Compared with the conventional manufacturing method EDS IgY, preparation process is much more effective: the number of serum producers is significantly lower, whereas the yield of the product is higher. Reduction of the volume of the immunogens, immunization cycle, and number of injections is also an advantage of this manufacturing method.