The Sakarya River basin is one of the largest basins in Turkey, and encompasses the Kocaeli, Düzce, Sakarya, Bursa, Bilecik, Bolu, Kütahya, Eskişehir, Ankara, Afyon, and Konya provinces. In this study, the water quality status of the basin was investigated using 18 diatom indices, calculated in Omnidia software. For this purpose, a total of 46 stations were surveyed in the rivers and streams of the basin in May 2018. As a result, 41 of 195 diatom taxa were found to be the most frequent (>10% share in assemblage). According to Detrented Correspondence Analysis (DCA), three subgroups were described as the spring section, Ankara and Polatlı section, and lowland section. The river basin quality was evaluated as moderate or lower quality status, while only a few sites had good status. The diatom index scores showed that the Descy’s Index (DES), Pampean Diatom Index (IDP), Artois-Picardie Diatom Index (IDAP), and Specific Pollution Sensitivity Index (IPS) appear best suited to water quality assessment in this area, showing the largest number of significantly important correlation with environmental variables.