We investigated the risk of maternal contamination in antenatal DNA diagnosis after second- and third-trimester transabdominal placental biopsy. For this purpose, we compared the restriction fragment length polymorphism (RFLP) patterns of 11 chorionic villus DNA samples after late chorionic villus sampling (CVS) with those of the corresponding maternal DNA. Ten of 11 aspirated tissue samples were not separated from maternal contamination before DNA extraction. All 11 mother-embryo pairs were informative for analysis of maternal contamination, ie, the mother showed one RFLP allele not present in the embryo. In none of the 11 cases did the fetal DNA show maternal contamination after molecular hybridization, although ten samples were contaminated with maternal tissue macroscopically and microscopically. Despite some maternal tissue admixture, the risk of contamination seems to be lower in the second- and third-trimester CVS than in first-trimester CVS, based on previous reports and our own experiences. This is most likely due to the anatomically closer contact of villi and decidua in the first trimester of pregnancy.