Lucilia eximia (Diptera: Calliphoridae) is a species of forensic importance for its association with decomposing bodies in Antioquia and other regions of Colombia. The implementation of methods to allow the rapid and accurate identification of this species is a matter important in forensic entomology. Currently, the mitochondrial DNA sequences are considered to complement classical taxonomy and are used in the characterization of morphologically similar or indistinguishable species as in the case of forensic importance fies. We evaluated the variability of a mitochondrial region (Cytb -RNAt-Ser-ND1) of 480 bp that was amplifed by PCR, from L. eximia larvae on third stage from Medellin and two other localities. The sequence differences evaluated as nucleotidic differences and genetic distances permitted the separation of individuals from the species of interest when compared with morphologically similar and related species. Furthermore, using a N.J analysis (K2p) the haplotype groupings were related to the geographical origin of individuals from L. eximia.