Technical Abstract: Comparative sequence analysis of six independent chicken and turkey parvovirus nonstructural (NS) genes revealed specific genomic regions with 100% nucleotide sequence identity. A PCR assay with primers targeting these conserved genome sequences proved to be highly specific and sensitive to detect parvoviruses in experimentally infected chickens. In a nationwide survey a total of 138 field enteric samples from poultry flocks were tested for parvovirus presence by PCR. Of the tested chicken samples that were collected in 54 farms, 77% showed the presence of parvovirus while 78% of the turkey samples that were received from 29 farms were parvovirus positive. For the first time, our data clearly demonstrate that parvoviruses are widely distributed in commercial poultry flocks in the US. The high prevalence of parvovirus infection in birds from enteric disease affected flocks suggests a potential role of these viruses in the etiology of enteric disease of poultry. Phylogenetic analyses comparing NS gene segments showed that most of the chicken and turkey parvovirus isolates formed separate phylogenetic groups. These findings suggest that the chicken and turkey parvoviruses might have diverged from a common ancestor and subsequently went through a host specific adaptation. The high level of sequence identity between the chicken and turkey parvoviruses indicates that these parvoviruses may cause similar diseases in their respective hosts.