[Objective] The aim of this study is to analyze the microbial diversity in the sediment of Jingshan hot spring. [Method] We extracted environment DNA from the sediment and then amplified the 16S rRNA genes of prokarotype and internal transcribed spacers(ITS) DNA of eukarotype by PCR with specific conservative primers. The PCR products were ligated to the T vectors and transformed into Escherichia coli to construct libraries. The libraries were analyzed by amplified ribosomal DNA restriction analysis (ARDRA) approach. The clones of dominant ARDRA patterns were selected to sequence. The sequence similarities were analyzed by using the BLAST programs for searching the GenBank DNA data-bases. [Results] We identified 14 dominant operational taxonomic units (OTUs) from prokarotype G. group. The Phy-logenetic tree shows that 7 of them (JS-GU20, JS-GU1, JS-G37, JS-G29, JS-GM2, JS-G42, JS-G41) belong to Bdellovi-brio. They had the highest similarity to Bacteriovorax sp. NE1 (EF092445) and Bdellovibrio sp. JS5 (AF084859), 96% and 99% respectively. Four major OTUs of prokarotype X group belong to Cyanobacterium. One of them, named JS-X2, has a similarity of 95% with an uncultured Cyanobacterium (L35331) which was gained from a hot spring of the Yellow-stone National Park and has a similarity of 89% with Thermosynechococcus elongatus BP-1 (47118315). Most OTUs of eukaryotic group have 90% sequences similarity to that of Penicillium sp.. [Conclusion] The results showed that the mi-crobial diversity in the sediment of Jinshan hot spring is rich. And the result of several clones’ similarity to Bacteriovorax sp. showed that there might be a new group of thermophilic Bacteriovorax sp. in the hot spring sediment.