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Background  In Benin, very few studies have been done on the genetics of  Plasmodium falciparum  and the resistance markers of anti-malarial drugs, while malaria treatment policy changed in 2004. Chloroquine (CQ) and sulphadoxine pyrimethamine (SP) have been removed and replaced by artemisinin-combination therapy (ACT). The objective of this study was to determine the genetic diversity of  P. falciparum  and the prevalence of  P. falciparum  molecular markers that are associated with resistance to CQ and SP in northern Benin seven years after the new policy was instituted.    Methods  The study was conducted in northern Benin, a region characterized by a seasonal malaria transmission. Blood samples were collected in 2012 from children presenting with asymptomatic  P. falciparum  infections. Samples collected in filter paper were genotyped by primary and nested PCR in block 2 of  msp-1  and block 3 of  msp-2  to analyse the diversity of  P. falciparum.  The prevalence of critical point mutations in the genes of  Pfcrt  (codon 76),  Pfmdr1  (codon 86),  Pfdhfr  (codons, 51, 59 and 108) and  Pfdhps  (codons 437, 540) was examined in parasite isolates by mutation-specific restriction enzyme digestion.    Results  Genotyping of 195 isolates from asymptomatic children showed 34  msp-1  and 38  msp-2  genotypes. The multiplicity of infection was 4.51 ± 0.35 for  msp-1  and 4.84 ± 0.30 for  msp-2.  Only the codon 51 of  Pfdhfr  and codon 437 of  Pfdhps  showed a high mutation rate: I51: 64.4% (57.3; 71.2); G437: 47.4% (40.2; 54.7), respectively. The prevalence of  Pfdhfr  triple mutant IRN (I51, R59 and N108) was 1.5% (0.3; 3.9), and  Pfdhfr/Pfdhps  quadruple mutant IRNG ( Pfdhfr I51, R59, N108, and  Pfdhps G437): 0. 5% (0; 2.5). No mutation was found with codon 540 of  Pfdhps . Analysis of mutation according to age (younger or older than ten years) showed similar frequencies in each category without significant difference between the two groups.    Conclusions  This study showed a high diversity of  P. falciparum  in northern Benin with a very low prevalence of resistance markers to CQ and SP that dramatically contrasted with the pattern observed in southern Benin. No influence of age on genetic diversity of  P. falciparum  and on distribution of the mutations was observed.

malaria journal

Low prevalence of the molecular markers of Plasmodium falciparum resistance to chloroquine and sulphadoxine/pyrimethamine in asymptomatic children in Northern Benin