Molecular determinants of sulfadoxine-pyrimethamine resistance in Plasmodium falciparum in Nigeria and the regional emergence of dhps 431V

Mary C Oguike, Catherine O Falade, Elvis Shu, Izehiuwa G Enato, Ismaila Watila, Ebenezer S Baba, Jane Bruce, Jayne Webster, Prudence Hamade, Sylvia Meek, Daniel Chandramohan, Colin J Sutherland, David Warhurst, Cally Roper, Mary C Oguike, Catherine O Falade, Elvis Shu, Izehiuwa G Enato, Ismaila Watila, Ebenezer S Baba, Jane Bruce, Jayne Webster, Prudence Hamade, Sylvia Meek, Daniel Chandramohan, Colin J Sutherland, David Warhurst, Cally Roper

Abstract

There are few published reports of mutations in dihydropteroate synthetase (dhps) and dihydrofolate reductase (dhfr) genes in P. falciparum populations in Nigeria, but one previous study has recorded a novel dhps mutation at codon 431 among infections imported to the United Kingdom from Nigeria. To assess how widespread this mutation is among parasites in different parts of the country and consequently fill the gap in sulfadoxine-pyrimethamine (SP) resistance data in Nigeria, we retrospectively analysed 1000 filter paper blood spots collected in surveys of pregnant women and children with uncomplicated falciparum malaria between 2003 and 2015 from four sites in the south and north. Genomic DNA was extracted from filter paper blood spots and placental impressions. Point mutations at codons 16, 50, 51, 59, 108, 140 and 164 of the dhfr gene and codons 431, 436, 437, 540, 581 and 613 of the dhps gene were evaluated by nested PCR amplification followed by direct sequencing. The distribution of the dhps-431V mutation was widespread throughout Nigeria with the highest prevalence in Enugu (46%). In Ibadan where we had sequential sampling, its prevalence increased from 0% to 6.5% between 2003 and 2008. Although there were various combinations of dhps mutations with 431V, the combination 431V + 436A + 437G+581G+613S was the most common. All these observations support the view that dhps-431V is on the increase. In addition, P. falciparum DHPS crystal structure modelling shows that the change from Isoleucine to Valine (dhps-431V) could alter the effects of both S436A/F and A437G, which closely follow the 2nd β-strand. Consequently, it is now a research priority to assess the implications of dhps-VAGKGS mutant haplotype on continuing use of SP in seasonal malaria chemoprevention (SMC) and intermittent preventive treatment in pregnancy (IPTp). Our data also provides surveillance data for SP resistance markers in Nigeria between 2003 and 2015.

Keywords: Nigeria; Sulfadoxine-pyrimethamine; dhfr; dhps; mutations.

Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Figures

Graphical abstract
Graphical abstract
Fig. 1
Fig. 1
Prevalence of dhps 431 mutation haplotypes between 2003 and 2014 in Nigeria. A map of Nigeria showing the study locations and the prevalence of the various combinations of the dhps-431V haplotypes between 2003 and 2014/2015. The prevalence of dhps-431V haplotypes in Yaounde, Cameroon (2015) is also shown.
Fig. 2
Fig. 2
Full model PfDHPS showing H-bonded parallel β-strands 1 and 2. At the C-terminus of β-2 is loop 2 containing substrate-binding mutable residues Ser436 and Ala437.
Fig. 3
Fig. 3
Clustal-0 alignment of DHPS sequences to locate structural features. This is largely from Pemble et al. (2010) but sequences used, apart from P. falciparum, are from FASTA Texts published with the crystal structures.

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