Chrysomya Robineau-Desvoidy is one of the most forensically important blow fly genera. The disadvantages of morphological taxonomy have resulted in numerous synonyms and caused problems in the species identification of this genus. In recent years, DNA barcoding (partial sequence of the mitochondrial cytochrome c oxidase subunit I (mtCOI) gene) has become a useful and popular tool for overcoming these difficulties. However, relevant studies generally focus on forensically-related species in specific regions. The insufficient sampling within specific genera in these studies limits their application in uninvestigated areas and precludes further insights into the actual availability of DNA barcoding for species identification. For this, the majority of currently available barcoding sequences of Chrysomya species are involved in a molecular analysis using different species delimitation methods. The results indicate that only up to 20 of 26 Chrysomya species can be distinctly separated from the others. With the integration of other information and a secondary genetic marker, the remaining ambiguous Chrysomya species are expected to be correctly separated.