The Papaya ringspot virus (PRSV), a member of the potyvirus that is transmitted by aphids within the Potyviridae family, is the main limiting factor for papaya (Carica papaya L.) and Cucurbits worldwide and causes losses of up to 100%. In this study, we conducted research on the genetic diversity of PRSV isolates collected from two locations in the department of Norte de Santander, Colombia. The analysis was performed by comparing the nucleotide sequences of the region that encode the coat protein (CP) of nine PRSV isolates from the Villa del Rosario location and 12 isolates from the Campo Hermoso location. The analysis included three sequences of the CP of PRSV isolates reported in the Colombian departments of Arauca and Valle del Cauca. The bioinformatic analysis suggested that the PRSV isolates from the locations in Norte de Santander were different from each other, grouping into different phylogenetic groups. Anexistence of recombination events in the coat protein was observed. This is the first study of PRSV genetic variability that has been conducted at the local level in Colombia.