Increased prevalence of the Plasmodium falciparum Pfmdr1 86N genotype among field isolates from Franceville, Gabon after replacement of chloroquine by artemether–lumefantrine and artesunate–mefloquine

Abstract

Despite global antimalarial measures, Plasmodium falciparum malaria remains a major public health problem. WHO has recommended the use of arteminisin-based combination therapy to limit the emergence of antimalarial drug resistance. However, ACT treatment failures have been linked to the selection of the wild types 86N genotype of P. falciparum multidrug resistance 1 (Pfmdr1) and the 76K genotype of P. falciparum chloroquine resistance (Pfcrt) genes. The aim of this study was to investigate the molecular impact of widespread implementation of artemether–lumefantrine and artesunate–mefloquine on local parasite population in Franceville, Gabon.We analyzed 230 pediatric field isolates (96 from 2004 and 134 from 2009). Routine hematological parameters were collected. Pfmdr1 codons 86 and 1246 and Pfcrt codon 76 were genotyped using PCR–RFLP and the prevalence of the genotypes was compared.The children's mean age did not differ between 2004 and 2009 (respectively 31.8 (6–84) months vs 38.6 (6–84) months, p=0.32), and neither did mean parasitemia [16,750 (1000–96,234) and 14,587 (1093–83,941) parasites/μL, respectively (p=0.21)]. The mean hemoglobin level was higher in 2009 than in 2004 (11.0±2.4 vs 7.8±2.0g/dL, respectively; p=0.04). More interesting, the prevalence of Pfmdr1 wild type 86N increased from 15.6% (n=15/96) in 2004 to 31.3% (n=42/134) in 2009 (p=0.007). A significant increase combining pure and mixed genotypes (86N+86N/Y) was also found between 2004 and 2009 (p=0.02), while the prevalence of genotypes Pfmdr1 1246D, Pfcrt wild type 76T and all mixed genotypes (Pfmdr1 86N/Y and 1246D/Y, and 76K/T) remained stable. The complexity of isolates was high (around 2.9 and 2.4) and the FC27 allele of Pfmsp2 was more prevalent.These findings show a substantial benefice of artemether–lumefantrine and artesunate–mefloquine and of new control measures. The selection, in the general population, of wild type Pfmdr1 86N, which is associated with antiplasmodial resistance against some drugs, has been induced underlining the need for molecular surveillance of the impact of ACT on antimalarial resistance.