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Background  A reversal of chloroquine (CQ) resistance following a period of withdrawal has raised the possibility of its re-introduction. This study evaluated the current prevalence of  Pfcrt  and  Pfmdr1  alleles in  Plasmodium falciparum  isolates, 11 years after CQ withdrawal in Southeast Nigeria.    Methods  Filter-paper blood samples were collected from 725 non-febrile individuals, comprising 250 children (≤ 12 years), 250 pregnant women and 225 other adults, between October 2014 and February 2015 in Nnewi town, Southeast Nigeria. Nested PCR followed by direct sequencing was employed for the genotyping of  Pfcrt  and  Pfmdr1  genes.    Results  A total of 103 parasites-positive samples were recovered, comprising of 48 (19.20%) among children, 20 (20.00%) among pregnant women and 35 (15.50%) among other adults cohort. The frequency of the mutant genotype of  Pfcrt  76T, 75E and 74I was 94.50% each. Parasite isolates from children had a frequency of 100% for mutant alleles in all  Pfcrt  codons while isolates from pregnant women and other adults had a frequency of 91% each in all codons. Haplotype distribution of  pfcrt  gene were 5.45, 0.00 and 76.37% for CVMNK, SVMNT and CVIET, respectively. For  Pfmdr1  gene, the frequency of 86Y, 184F and 1246Y mutant alleles were 8.54, 29.27 and 3.66%, respectively. Amongst the  Pfmdr1  haplotypes analysed, NFD had the highest frequency of 24.4%, followed by YFD at 6.10%. NYF and NYY occurred the least (1.20%).    Conclusion  The high level of  Pfcrt  mutations is suggestive of a sustained CQ pressure on  P. falciparum  isolates in the study area, despite the change of first line treatment from CQ to artemisinin combination therapy for 11 years. A new strategy to ensure the complete withdrawal of CQ from the country is recommended.

malaria journal

Analysis of Plasmodium falciparum Pfcrt and Pfmdr1 genes in parasite isolates from asymptomatic individuals in Southeast Nigeria 11 years after withdrawal of chloroquine