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01.12.2012 | Research | Ausgabe 1/2012 Open Access

Malaria Journal 1/2012

Temporal trends of sulphadoxine-pyrimethamine (SP) drug-resistance molecular markers in Plasmodium falciparum parasites from pregnant women in western Kenya

Malaria Journal > Ausgabe 1/2012
Nnaemeka C Iriemenam, Monica Shah, Wangeci Gatei, Anna M van Eijk, John Ayisi, Simon Kariuki, Jodi Vanden Eng, Simon O Owino, Ashima A Lal, Yusuf O Omosun, Kephas Otieno, Meghna Desai, Feiko O ter Kuile, Bernard Nahlen, Julie Moore, Mary J Hamel, Peter Ouma, Laurence Slutsker, Ya Ping Shi
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​1475-2875-11-134) contains supplementary material, which is available to authorized users.

Competing interests

The authors declare that they have no competing interests.

Author’s contributions

NCI, MS, WG, SK, AAL and YOO carried out genotyping work for the current study. AME, JA, FOK, BN (first period), JM, SOO (second period), MD, MJH, PO, and LS (third period) implemented the observational malaria in pregnancy studies from 1996 to 2009 including enrolment of patients and collection of clinical and epidemiological data and samples. YPS conceived, designed and supervised the current study. NCI, MS and JVE did the statistical analysis. NCI, MS and YPS wrote the paper. All authors contributed to the interpretation of results and critical discussion of the conclusion and approved the final manuscript.



Resistance to sulphadoxine-pyrimethamine (SP) in Plasmodium falciparum parasites is associated with mutations in the dihydrofolate reductase (dhfr) and dihydropteroate synthase (dhps) genes and has spread worldwide. SP remains the recommended drug for intermittent preventive treatment for malaria in pregnancy (IPTp) and information on population prevalence of the SP resistance molecular markers in pregnant women is limited.


Temporal trends of SP resistance molecular markers were investigated in 489 parasite samples collected from pregnant women at delivery from three different observational studies between 1996 and 2009 in Kenya, where SP was adopted for both IPTp and case treatment policies in 1998. Using real-time polymerase chain reaction, pyrosequencing and direct sequencing, 10 single-nucleotide polymorphisms (SNPs) of SP resistance molecular markers were assayed.


The prevalence of quintuple mutant (dhfr N51I/C59R/S108N and dhps A437G/K540E combined genotype) increased from 7 % in the first study (1996–2000) to 88 % in the third study (2008–2009). When further stratified by sample collection year and adoption of IPTp policy, the prevalence of the quintuple mutant increased from 2.4 % in 1998 to 44.4 % three years after IPTp policy adoption, seemingly in parallel with the increase in percentage of SP use in pregnancy. However, in the 1996–2000 study, more mutations in the combined dhfr/dhps genotype were associated with SP use during pregnancy only in univariable analysis and no associations were detected in the 2002–2008 and 2008–2009 studies. In addition, in the 2008–2009 study, 5.3 % of the parasite samples carried the dhps triple mutant (A437G/K540E/A581G). There were no differences in the prevalence of SP mutant genotypes between the parasite samples from HIV + and HIV- women over time and between paired peripheral and placental samples.


There was a significant increase in dhfr/dhps quintuple mutant and the emergence of new genotype containing dhps 581 in the parasites from pregnant women in western Kenya over 13 years. IPTp adoption and SP use in pregnancy only played a minor role in the increased drug-resistant parasites in the pregnant women over time. Most likely, other major factors, such as the high prevalence of resistant parasites selected by the use of SP for case management in large non-pregnant population, might have contributed to the temporally increased prevalence of SP resistant parasites in pregnant women. Further investigations are needed to determine the linkage between SP drug resistance markers and efficacy of IPTp-SP.
Additional file 1: Statistical procedure. The file described the statistical methods used for the descriptive and association analysis. (DOC 47 KB)
Additional file 2: Proportion of SP drug resistant genotypes by HIV status and by study period. A, dhfr genotypes. B, dhps genotypes. C, combined dhfr/dhps genotypes. The figures described the prevalence of dhfr, dhps and the combined dhfr/dhps genotypes between HIV + and HIV- women. (TIFF 361 KB)
Authors’ original file for figure 1
Authors’ original file for figure 2
Authors’ original file for figure 3
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Authors’ original file for figure 5
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