In order to screen out lymphocystis disease resistance-related SSR markers in Chinese flounder population, 102 individuals were used(56 disease susceptible and 46 disease resistant individuals) in the present study.Firstly, two gene pools were constructed using 15 susceptible individuals and 15 resistant individuals, respectively.The two gene pools were scanned using 178 pairs of microsatellite primers.Secondly, the differential bands amplified in gene pools were verified using 30 individuals which were used to construct the gene pools for the first time.Lastly, a second verification for 102 individuals was performed to verify the difference significance of the differential bands identified in the first verification.Results of BSA analysis demonstrated that some differential bands were amplified using four pairs of primers(scaffold440_22585, scaffold826_5003, scaffold703_4284 and scaffold185_597).The first verification indicated that the differential bands are significantly different in the SSR markers scaffold826_5003(P=0.023) and scaffold185_597(P_178bp=0.028, P_173bp=0.009).And in the second verification, the difference was extremely significant in the totally 102 individuals with the SSR marker scaffold185_597, the frequency of differential bands in disease resistant individuals and disease susceptible individuals was 60.9% and 14.3%, respectively(P=0.001<0.01).The differential bands amplified from ten disease resistant individuals using primer scaffold185_597 were cloned and sequenced.Sequence alignment by BLAST confirmed that the bands were fragments of microsatellite marker scaffold185_597 published by lab for aquatic genomics and cell engineering in Yellow Sea Fisheries Research Institute, homology of which was up to 96%.The present study has shown that the microsatellite marker scaffold185_597 may be associated with lymphocystis disease resistance in Japanese flounder(Paralichthys olivaceus), providing some basis for the molecular marker-assisted selection in Chinese flounder.