To evaluate the in vitro chemosensitivity to Atovaquone (ATQ) and analyze polymorphism of Pfcytb gene of Plasmodium falciparum isolates from Abidjan. Fifty seven isolates of P. falciparum collected between 2010 and 2012 from the outskirts of the city of Abidjan. Were used for testing in vitro susceptibility according to the microtest optical version of the World Health Organization (WHO). Polymerase Chain Reaction (PCR) was used to determine mutation in the Pfcytb gene. The analysis of the relationship between the observed mutations and chemosensitivity of isolates was made using Cohen's kappa test. The geometric mean IC50 of atovaquone is 1,645 mM. All 57 isolates were sensitive to atovaquone. The 29 isolates of P. falciparum sequenced showed no mutation of Pcytb gene at codon Y268. No other mutation was observed. Our study showed no correlation between phenotypic and molecular data (K< 0). This study shows that the combination atovaquone / proguanil can be one of the drugs of choice for the prophylaxis of multidrug-resistant falciparum malaria recommended by the World Health Organization (WHO).