This in-silico study aimed to determine the phylogenetic relationship of Listeria monocytogenes 16S ribosomal RNA and hly virulence gene sequences. The sequences were obtained from the National Center for Biotechnology Information (NCBI). A total of 20 strains obtained from NCBI were randomly selected by country from different continents and considered the source of isolation, temperature/season, and period of time when the bacteria were isolated. Result showed that there were random evolutionary occurrences formed in /6S rRNA gene sequences and Aly gene sequences. Analysis suggested that there is difference between the constructed phylogenetic trees of /6S rRNA and hly gene sequences in which /6S rRNA genes formed three groups; two major groups and one group containing single evolutionary lineage. However, hiy gene sequences formed three groups and one out-group; one major group, another group with single evolutionary lineage and another group forming monophyletic clade and single out-group, and the evolution of L. monocytogenes sequences are not affected by their different conditions (country, source, temperature/season and year when the bacterium was isolated). Keywords: Listeria monocytogenes, 16S rRNA gene, hily gene, Phylogenetic tree.