The Sanshenjiang gold deposit in southeastern Guizhou Province, China, is hosted by the Neoproterozoic metasedimentary rocks which experienced low-grade greenschist facies metamorphism. Gold mineralization occurs mainly in the ribbon chiltern slate of the first member of the Longli Formation and is controlled by both strata and faults. Ore bodies are characterized by abundant quartz-arsenopyrite-gold-pyrite-bedding veins, veinlets and small lenses within the shear zone. In this study, trace element and REE geochemistry was analyzed to constrain the origin and genesis of this deposit. The trace element signatures of wall rocks and veins display a basically similar tendency in the spider diagram, showing the genetic relationship. The values of Co/Ni, Y/Ho, Hf/Sm, Nb/La and Th/La reflect that the hydrothermal fluids of this deposit were derived from the mixture of multiple sources with marked enrichment of Cl and moderate to high temperature. There is a broad similarity in the chondrite-normalized patterns and REE fractionation between wall rocks and ore bodies, possibly reflecting their similar origin. Based on the difference in δCe and δEu, quartz veins and lenses can be subdivided into weakly negative Ce-anomalies (δCe=0.81 to 1.06) with slight Eu anomalies (δEu=0.81 to 1.06) type and the significant positive Ce-anomalies (δCe=1.13 to 1.97) with moderate negative Eu-anomalies type, probably suggesting physical-chemical changes in the evolution process of ore-forming fluids from the early to late stage. It can be concluded that the ore-forming process may have experienced three stages: formation of the original ore source bed, regional metamorphism and gold mineralization, on the basis of trace element and REE analysis and field observation. The Sanshenjiang gold deposit in southeastern Guizhou Province, China, is hosted by the Neoproterozoic metasedimentary rocks which are low-grade greenschist facies metamorphism. Gold mineralization occurs mainly in the ribbon chiltern slate of the first member of the Longli Formation and is controlled by both strata and faults. Ore bodies are characterized by abundant quartz-arsenopyrite-gold-pyrite-bedding veins, veinlets and small lenses within the shear zone. In this study, trace element and REE geochemistry was analyzed to constrain the origin and genesis of this deposit. The values ​​of Co / Ni, Y / Ho, Hf / Sm, Nb / La and Th / La reflect that the hydrothermal fluids of this deposit were derived from the mixture of multiple sources with marked enrichment of Cl and moderate to high temperature. There is a broad similarity in the chondrite-n Based on the difference in δCe and δEu, quartz veins and lenses can be subdivided into weakly negative Ce-anomalies (δCe = 0.81 to 1.06) with slight Eu anomalies (δEu = 0.81 to 1.06) type and the significant positive Ce-anomalies (δCe = 1.13 to 1.97) with moderate negative Eu-anomalies type, probably suggesting physical-chemical changes in the evolution process of ore-forming fluids from the early to late stage. It can be concluded that the ore-forming process may have experienced three stages: formation of the original ore source bed, regional metamorphism and gold mineralization, on the basis of trace element and REE analysis and field observation.