MOLECULAR EPIDEMIOLOGY OF MALARIA IN CAMEROON. XVII. BASELINE MONITORING OF ATOVAQUONE-RESISTANT PLASMODIUM FALCIPARUM BY IN VITRO DRUG ASSAYS AND CYTOCHROME B GENE SEQUENCE ANALYSIS

Abstract

Atovaquone is a new broad-spectrum antiprotozoal drug with high in vitro activity against multidrug-resistant Plasmodium falciparum. Its specific action against protozoans is based on the inhibition of the parasite cytochrome bc1 complex of the mitochondrial electron transport system. Protozoans may develop atovaquone resistance by the selection of a mutant cytochrome b gene. With the increasing availability of atovaquone-proguanil combination for prophylaxis and treatment of malarial infections, it is necessary to establish baseline data on atovaquone sensitivity before the drug is introduced massively in an endemic region. For this purpose, the activity of atovaquone was assessed indirectly by in vitro drug sensitivity assays with several serum substitutes and DNA sequencing of the cytochrome b gene. Using the standard in vitro assay procedures with 10% human serum, the geometric mean 50% inhibitory concentration (IC50) for atovaquone was calculated to be 1.15 nM (range = 0.460-4.17 nM), while the use of 10% fetal calf serum resulted in lower IC50s (geometric mean = 0.575, range = 0.266-2.20 nM). The use of Albumax, a lipid-enriched bovine albumin, over the same concentration range (0.25-16 nM) showed poor results. None of the 37 isolates with an atovaquone IC50 < 4.17 nM displayed any mutation. Further monitoring of atovaquone-resistant P. falciparum is warranted for the rational use of this new antimalarial drug.