The aims of this study were to analyze the physico-chemical factors and the characteristics of epilithic diatom community from 15 sites of the Naeseongcheon and tributaries located in the upper region of the Nakdong river from May to October 2016. The biological water quality was assessed using DAIpo and TDI. A total of 163 diatom taxa were identified with 2 orders, 3 suborders, 9 families, 35 genera, 145 species, 16 varieties and 2 forms. Cocconeis placentula var. lineata appeared at every examined sites. Achnanthes lanceolata, Nitzschia fonticola, Nitzschia inconspicua and Reimeria sinuata were common taxa of the Naeseongcheon. Nitzschia inconspicua and Achnanthes minutissima were major dominant species. As a result of the CCA, Electrical conductivity and total nitrogen concentration were important factors determining the diatom species composition. In the result of the biological assessment using DAIpo, the Naeseongcheon was rated at class B with an average of 62.38. In the result of assessment using TDI, the Naeseongcheon was rated at class C with an average of 66.12.