The bacteria belonging to Raoultella and Klebsiella genera are opportunistic pathogens for which there is no consensus about a unique internationally taxonomic system. In this study, we suggest a molecular phylogeny based on 16S (rDNA 16S) ribosomal gene and the beta subunit RNA polymerase (rpoB) encoding gene of Klebsiella and Raoultella in order to set up evolutionary relationships among these genera. Results showed a cluster with similar biochemical and taxonomy characteristics in agreement to Genbank description, and a tree bifurcation which confirms the Klebsiella and Raoultella genera separation. Moreover, we verified the polyphyletic character of K. aerogenesby rDNA 16S and particularly the clustering for both R. terrigena and K. oxytoca based on rpoB gene. The evolutionary relationships recognition was obtained by comparison among the corresponding trees for both genes unraveling significant changes at taxonomic level and stressing the importance of multilocus analysis approach. These studies are useful for tracking the microbial genotype stability over time and space scale; as well as on improving taxonomic annotations (taxa descriptions and genetic subdivisions) and evaluation of genetic diversity interms of virulence and drug resistance.