Assessment of Plasmodium falciparum drug resistance molecular markers from the Blue Nile State, Southeast Sudan

Abstract

Background: Plasmodium falciparum malaria is a public health problem worldwide. Malaria treatment policy has faced periodic changes due to emergence of drug resistant parasites. In Sudan chloroquine has been replaced by artesunate and sulfadoxine/pyrimethamine (AS/SP) in 2005 and to artemether-lumefantrine (AL) in 2017, due to the development of drug resistance. Different molecular markers have been used to monitor the status of drug resistant P. falciparum. This study aimed to determine the frequency of malaria drug resistance molecular markers in Southeast Sudan. Methods:The samples of this study were day zero dried blood spot samples collected from efficacy studies in the Blue Nile State from November 2015 to January 2016. A total of 130 samples were amplified and sequenced using illumina Miseq platform. The molecular markers included were Pfcrt, Pfmdr1, Pfdhfr, Pfdhps, Pfk13, exonuclease and artemisinin resistant (ART-R) genetic background (Pfmdr2, ferroredoxine, Pfcrt and Pfarps10). Results:Resistance markers for chloroquine were detected in 25.8% of the samples as mutant haplotype Pfcrt 72-76 CVIET and 21.7% Pfmdr1 86Y. Pfdhfr mutations were detected in codons 51, 59 and 108. The ICNI double-mutant haplotype was the most prevalent (69%). Pfdhps mutations were detected in codons 436, 437, 540, 581 and 613. The SGEGA triple-mutant haplotype was the most prevalent (43%). In Pfdhfr/Pfdhps combined mutation, quintuple mutation ICNI/SGEGA is the most frequent one (29%). Six of the seven treatment failure samples had quintuple mutation and the seventh was quadruple. This was significantly higher from the adequately responsive group (P < 0.01). Pfk13 novel mutations were found in 7 (8.8%) samples, which were not linked to artemisinin resistance. Mutations in ART-R genetic background genes ranged from zero to 7%. Exonuclease mutation was not detected.

Efficacy of Dihydroartemisinin-piperaquine Versus Artemether-lumefantrine for the Treatment of Uncomplicated Plasmodium Falciparum Malaria Among Children in Africa: a Systematic Review and Meta-analysis of Randomized Control Trials

“…A target dose (range) of 4 (2-10) mg/kg bw per day dihydroartemisinin and 18 (16-27) mg/kg bw per day piperaquine given once a day for 3 days for children weighing ≥ 25 kg. The target doses and ranges for children weighing < 25 kg are 4 (2.5-10) mg/kg bw per day dihydroartemisinin and 24 (20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32) mg/kg bw per day piperaquine once a day for 3 days.…”

“…Resistance to ACT in Southeast Asia is becoming the highest concern [18]. There are a few reports on artemisinin resistance mediated by mutations kelch13 (K13) gene in Greater Mekong Sub-region [19], Sudan [20], higher prevalence (42%) in Myanmar [21], and low frequency of kelch13 (K13) gene mutation in 18 Sub-Saharan African countries [22,23]. In addition, over the past ten years a decline in parasitological response in Nigeria [24], decrease in PCR corrected therapeutic e cacy of ACT below 80% in Burkina Faso [25] have been noticed, Moreover, increase in copy number of plasmepsin genes associated with decrease in effectiveness of piperaquine has been arisen in South East Asia [26][27][28].…”

<p>Trends in Molecular Markers Associated with Resistance to Sulfadoxine-Pyrimethamine (SP) Among <em>Plasmodium falciparum</em> Isolates on Bioko Island, Equatorial Guinea: 2011–2017</p>

“…14 Previous report revealed that the combination of Pfdhfr-Pfdhps mutations significantly correlated with the treatment failure. 15 For the Pfdhfr gene, mutations N51I, C59R, S108N and I164L are associated with pyrimethamine resistance. Similarly, five mutations in Pfdhps (S436A, A437G, K540E, A581G and A613S/T) have been implicated in sulfadoxine resistance.…”

Antimalarial drug efficacy and resistance in malaria‐endemic countries in HANMAT‐PIAM_net countries of the Eastern Mediterranean Region 2016–2020: Clinical and genetic studies

“…Both mutations have recently been reported to be associated with delayed parasite clearance in Rwanda and Uganda, respectively [7, 8]. No validated or associated Pfk13 mutations were detected in Sudan in other studies conducted between 2015 and 2017, the same time period as the current studies [41–43]. However, none of these latter studies were conducted at the Gadaref and Sennar sites where Pfk13 R622I mutation was detected.…”