Plutella xylostella, pointed out as the most dangerous destroyer of cabbage (Brassica olerarea), is a cosmopolitan species. In fact, owing to its large capacity of adaptation, P. xylestella colonizes any kind of area. This insect can bring about up to 90% of losses on cabbage farming. To control the insect, Senegalese farmers, very often, resort to high dose of chemical pesticides which are repetitively sprayed. The use of these chemical products gives rise to different varieties of resistant insects, which results in the emergence of different haplotypes between populations. It is in such a context that this study has been undertaken. Our objective is then to contribute to the knowledge of the genetic diversity of P. xylostella populations in Senegal. To hit the target, PCR- Sequencing method has been applied on samples from the five following localities: Diofior, Malika, Mboro, Santh Ndong, and Sebikotane. P. xylostella from Mboro is genetically different from the other populations. This fact could be due to a different way of using pesticides in this area vis-à-vis of the other localities. On the other hand, between populations of P. xylostella from Diofior, Malika, Santh Ndong, and Sebikotane, there is no significant genetic difference. In other respects, phylogenetic trees reveal the existence of two clades: one with individuals from Mboro and the other one with individuals from the other four localities. P. xylostella from Mboro distinguishes itself from other populations and then, can be regarded as a sub-population. Thus, the phylogenetic trees reveal the existence of two groups of P. xylostella in Senegal.