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Background  With the introduction of artemisinin-based combination therapy (ACT) in 2005, monitoring of anti-malarial drug efficacy, which includes the use of molecular tools to detect known genetic markers of parasite resistance, is important for first-hand information on the changes in parasite susceptibility to drugs in Ghana. This study investigated the  Plasmodium falciparum  multidrug resistance gene ( pfmdr1 ) copy number, mutations and the chloroquine resistance transporter gene ( pfcrt ) mutations in Ghanaian isolates collected in seven years to detect the trends in prevalence of mutations.    Methods  Archived filter paper blood blots collected from children aged below five years with uncomplicated malaria in 2003–2010 at sentinel sites were used. Using quantitative real-time polymerase chain reaction (qRT-PCR), 756 samples were assessed for  pfmdr1  gene copy number. PCR and restriction fragment length polymorphism (RFLP) were used to detect alleles of  pfmdr1  86 in 1,102 samples,  pfmdr1  184, 1034, 1042 and 1246 in 832 samples and  pfcrt  76 in 1,063 samples. Merozoite surface protein 2 ( msp2 ) genotyping was done to select monoclonal infections for copy number analysis.    Results  The percentage of isolates with increased  pfmdr1  copy number were 4, 27, 9, and 18% for 2003–04, 2005–06, 2007–08 and 2010, respectively. Significant increasing trends for prevalence of  pfmdr1  N86 (× 2  = 96.31, p <0.001) and  pfcrt  K76 (× 2  = 64.50, p <0.001) and decreasing trends in  pfmdr1  Y86 (× 2  = 38.52, p <0.001) and  pfcrt  T76 (× 2  = 43.49, p <0.001) were observed from 2003–2010. The  pfmdr1  F184 and Y184 prevalence showed an increasing and decreasing trends respectively but were not significant (× 2  = 7.39,p=0.060; × 2  = 7.49, p = 0.057 respectively). The  pfmdr1  N86-F184-D1246 haplotype, which is alleged to be selected by artemether-lumefantrine showed a significant increasing trend (× 2  = 20.75, p < 0.001).    Conclusion  Increased  pfmdr1  gene copy number was observed in the isolates analysed and this finding has implications for the use of ACT in the country although no resistance has been reported. The decreasing trend in the prevalence of chloroquine resistance markers after change of treatment policy presents the possibility for future introduction of chloroquine as prophylaxis for malaria risk groups such as children and pregnant women in Ghana.

malaria journal

Increased pfmdr1 gene copy number and the decline in pfcrt and pfmdr1 resistance alleles in Ghanaian Plasmodium falciparum isolates after the change of anti-malarial drug treatment policy