Filamentous bacteria overlaying ulcerated area on the body surface were observed in the wet-mout preparation from a moribund goldfish with saddle back appearance. The causative agent was identified as Flavobacterium columnrae, on the basis of biochemical test, species-specific polymerase chain reaction (PCR) and sequencing of 16S rDNA gene with the universal bacterial primers. Furthermore, the strain (ING-1) attributed to genomovar II in 16S rDNA PCR-restriction fragment length polymorphism (PCR-RFLP) and sequence analysis. In phylogenetic analysis, the strain ING-1, produced typical columnaris disease symptoms in rohu (Labeo rohita) fingerlings within 10 days. This is a new record about molecular detection and identification of Flavobacterium columnare, occurring naturally on a new host Carassius auratusin India.