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Background  The prevention of malaria faces with the repeated emergence of  Plasmodium falciparum  resistance to drugs, often involving point mutations of the target gene. In the pregnant woman, currently the WHO recommendation is the administration of an intermittent preventive treatment (IPTp) with sulphadoxine-pyrimethamine. Sulphadoxine-pyrimethamine (SP) resistance has increased for several years in Africa, stressing the need for alternative molecules. In this context, the first randomized clinical trial comparing the efficacy of SP and mefloquine for IPTp has been conducted recently in Benin. Using samples from this trial, the current study evaluated and quantified the prevalence of mutations on the  pfdhfr  and  pfdhps  genes as well as the copy number of the  pfmdr1  gene in parasites from  P. falciparum -infected pregnant women before first and second IPTp administration, and at delivery.    Methods  PCR-restriction fragment length polymorphism of polymorphic codons of the  pfdhfr  gene (51, 59, 108, and 164) was performed. The identification of mutations in three codons of the  pfdhps  gene (436, 437 and 540) was achieved by PCR and sequencing. Copy number quantification for  pfmdr1  gene was performed using real-time PCR.    Results  Results show a high prevalence rate of mutant parasites in women taking IPTp with sulphadoxine-pyrimethamine or mefloquine. The prevalence of triple and quadruple mutants was high before first drug regimen administration (79/93, 85%), and remained similar until delivery. Infection with mutant parasites was not correlated with low birth weight nor placental infection. In all samples, the copy number of  pfmdr1  gene was equal to one.    Conclusions  The clinical trial comparing SP and mefloquine efficacy during IPTp showed SP remained efficacious in preventing low birth weight. The present study shows a high prevalence of triple and quadruple mutations implicated in SP resistance. Although the  pfdhfr / pfdhps  triple and quadruple mutations were frequent, there was no evidence of correlation between these genotypes and the lack of efficacy of SP in the context of IPTp. Nevertheless, it is now obvious that SP will soon be compromised in whole Africa. Molecular markers have been recommended to monitor SP efficacy for IPTp, but given the current prevalence of mutant parasites their usefulness is questionable.

Molecular markers of resistance to sulphadoxine-pyrimethamine during intermittent preventive treatment of pregnant women in Benin