This study establishes a new method for paternity testing consisting of genotyping and PIA typing (parentally imprinted allele typing) of a VNTR locus 5' to the H19 imprinted gene. This procedure can be used to determine the parental origin of two alleles using differential DNA methylation. The PIA typing detects only the paternal allele of the two alleles detected by genotyping. The most suitable application for the PIA typing is paternity testing. While identification of the child’s paternal allele can usually be made in a paternity trio consisting of a mother, child, and alleged father, this is not possible when only the father is available for testing or when results depend on testing other second degree relatives such as grandparents or siblings of the alleged father. The PIA testing is also able to distinguish between two alleged fathers when the mother and the alleged father(s) share alleles such that both alleged fathers would be included. The method could also assist in resolving incest cases when multiple male relatives are suspected. In the case shown, the probability of paternity exclusion based on the PIA results was 82.6 percent, while the exclusion probability based on the genotyping results was 19.1 percent. This suggests that incorporation of the PIA testing into the conventional paternity test with STRs will give high resolution in paternity dispute cases. For forensic application, this method was applied to postmortem tissue DNAs from body parts. All provided only the paternal allele. The result suggests that the PIA typing provides useful information in forensic investigations. 4 figures, 1 table, 16 references