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<p>Abstract</p> <p>Background</p> <p><it>Plasmodium vivax</it> has traditionally been considered virtually absent from Western and Central Africa, due to the absence of the Duffy blood group in most of the population living in these areas. Recent reports, however, suggest the circulation of <it>P. vivax</it> in sub-Saharan Africa.</p> <p>Methods</p> <p>Giemsa/Field-stained smears from febrile patients recruited in five different cities (Goundam, Tombouctou, Gao, Bourem and Kidal) pertaining to three regions from Northern Mali were examined. Nested-PCR and DNA sequence analyses of selected samples were performed to fully confirm the presence of <it>P. vivax</it> infections.</p> <p>Results</p> <p>Results demonstrated the presence of <it>P. vivax</it> infections in close to 30% of the cases as detected by Giemsa/Field-stained smears and nested-PCR and DNA-sequence analyses of selected samples unequivocally confirmed the presence of <it>P. vivax</it>.</p> <p>Conclusions</p> <p>The diagnostics of this human malaria parasite should be taken into account in the context of malaria control and elimination efforts, not only in Mali, but also in sub-Saharan Africa.</p>