Abstract：Objective：To explore and analyze the changes in community structure of intestinal flora in patients with premature ovarian insufficiency (POI) with kidney deficiency and liver constraint syndrome based on 16S rDNA sequencing. Methods：A total of 60 cases of POI patients with kidney deficiency and liver constraint syndrome were included (POI group)， and 30 healthy volunteers were recruited(healthy control group). The characteristics of intestinal flora of POI patients with kidney deficiency and liver constraint syndrome and healthy volunteers were analyzed by high-throughput 16S rDNA sequencing，and comparative was conducted. Results： Compared with those in the healthy control group， at the phylum level，the relative abundance of bacteroides was increased in the POI group，and the relative abundance of firmicutes and actinomycetes was decreased； at the genus level， the relative abundance of provetella， bacteroides， and clostridium XIVa was increased in the POI group， and the relative abundance of fecal bacillus was decreased. Conclusion： There are differences in intestinal flora between POI patients and healthy groups，and the imbalance in the proportion of intestinal flora may cause disturbances in the internal environment of the organism，leading to the occurrence of POI.