Paternity index was analyzed using five microsatellite loci among Chinese egg-laying ducks (Anas platyrhynchos). Based on the paternity relationship that was identified by paternity index analysis, the estimated breeding value (EBV) was calculated using BLUP (best linear unbiased predictor) method. Body weight at first egg (BWF) is the only considered trait in this study. In total, 12 sires, 31 dams and 77 daughters were involved in the EBV calculation. The results demonstrated that five microsatellite loci's polymorphism information content (PIC) ranged from 0.795 in locus AY493338 to 0.957 in locus AY493264 with average 0.899; the parent-offspring relationships were built by these microsatellites' genotype, 12 families of half sibling and 2 families of full sibling were involved, and the relationship error is smaller than 10(-7). The EBV results suggest that the average EBV was significantly higher in females (average EBV is 10.234 and 0.1045 for mother and daughter, respectively) than males (average EBV is just -26.44). The EBV results on BWF were in good agreement with the principle of GH (growth hormone) expression in poultry. These results show that paternity analyses of Chinese egg-laying ducks were basically resolved using the five microsatellite loci selected. The paternity relationships can apply in Chinese egg-laying duck breeding to quicken the improvement of genetic progress.