Virus neutralization test (VNT) is considered to be the gold standard for arbovirus serology because of its high specificity and sensitivity. Its micromodification in 96-well plate with vital staining of cell cultures was developed in the National Reference Laboratory (NRL) for Arboviruses of the Czech Republic and is used for the detection of specific antibodies against various viruses, mainly arboviruses. The test procedure is described for the Tahyna virus micromodified neutralization assay in 96-well plate. Results of an anti-Tahyna antibody survey conducted among the population of northern Moravia using the VNT assay are presented. The overall anti-Tahyna seroprevalence among 1001 tested persons was 3.80%. The highest positivity rate was found in persons aged over 59 years (17.53%) comparing to children with the seroprevalence rates of 0.00% and 0.56% in the age groups 0-5 years and 6-14 years, respectively.