Identifying routes of invasion is a critical management strategy in controlling the spread of fish invasive species. This is challenging, however, in the absence of direct evidence. Therefore, indirect methodologies are used to infer possible invasion sources and routes, such as genetic and morphological data from populations from invasive ranges and putative source areas. During 2003 and 2014, 51 Perccottus glenii individuals were collected from three locations across Danube River basin: Silvia channel, Mures River, Romania (2009); Tiszafured, Hungary, Tisza River (2013) and Siret River, Romania (2014). The collected specimens were stored in 98% ethanol. A small fragment of dorsal muscle tissue was subject to genomic DNA extraction using phenol–chloroform-isoamyl alcohol (25:24:1) extraction protocol. Amplicons of 1150bp for mitochondrial gene cytochrome b (cytb) and 890bp mitochondrial control region were amplified by PCR using a new designed specific primers pair. The PCR products were purified using QIAquick PCR Purification Kit (Qiagen) and sequenced on a CEQ8000 Genetic Analysis System (Beckman Coulter). A cytb gene dataset was build using sequences from 51 Amur Sleepers from Europe combined with another 61 Asian haplotypes sequences from Xu et al. (2014) study and 4 from Thacker et al. (2005) study. Cytb gene dataset aligned with three outgroup species (Odontobutis obscura; O. potamophila and O. platycephala) was subject to jModelTest v.2, nucleotide substitution model being selected by Bayesian information criteria. The phylogenetic relationships within European and Asian Amur Sleepers were reconstructed using the ML method in PhyML v.3 software, and tested under 1000 bootstraps replications. The aim of the present study was to assess the native area and the invasivity of European Amur sleepers using two mitochondrial markers. In order to evaluate the invasiveness features, their genetic diversity across the native area in comparison with the genetic diversity within invasive areas, were computed.