Introduction: Anisakiasis is a human parasitic disease caused by the consumption of raw fish or shellfish containing larvae of the Anisakidae family. It is currently considered an emerging disease of public health interest.Objective: To identify the presence of larvae of the Anisakidae family in samples of frozen raw fish fillets intended for human consumption in markets in Medellín and its metropolitan area in Antioquia, Colombia.Materials and methods: A cross-sectional study was carried out, in which larvae of the Anisakidae family were detected and identified in frozen raw fish fillets from three representative markets in Medellín and its metropolitan area. A total of 384 ready for consumption fillets were analyzed (197 sawfish, 137 salmon, 37 tuna, and 13 hake), using the pressing and ultraviolet light method. Taxonomic keys were used to identify the collected parasites and to establish its genus. Conventional PCR and Sanger sequencing was performed to determine the species.Results: Four larvae were found in 4 of the 384 (1.04%) fillets (CI95% 1.04 ± 1.01%). The species of fish in which the larvae were found was sawfish (Scomberomorus spp.) and the genus and species of the larvae was established as Anisakis pegreffii.Conclusions: According to the study, the presence of Anisakis parasites in frozen raw fish fillets in the influence area is evident.