Prevalence of arps10, fd, pfmdr-2, pfcrt and pfkelch13 gene mutations in Plasmodium falciparum parasite population in Uganda
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Date
2022
Journal Title
Journal ISSN
Volume Title
Publisher
Plos one
Abstract
In Uganda, Artemether-Lumefantrine and Artesunate are recommended for uncomplicated
and severe malaria respectively, but are currently threatened by parasite resistance.
Genetic and epigenetic factors play a role in predisposing Plasmodium falciparum parasites
to acquiring Pfkelch13 (K13) mutations associated with delayed artemisinin parasite clearance
as reported in Southeast Asia. In this study, we report on the prevalence of mutations
in the K13, pfmdr-2 (P. falciparum multidrug resistance protein 2), fd (ferredoxin), pfcrt (P.
falciparum chloroquine resistance transporter), and arps10 (apicoplast ribosomal protein
S10) genes in Plasmodium falciparum parasites prior to (2005) and after (2013) introduction
of artemisinin combination therapies for malaria treatment in Uganda. A total of 200 P. falciparum
parasite DNA samples were screened. Parasite DNA was extracted using QIAamp
DNA mini kit (Qiagen, GmbH, Germany) procedure. The PCR products were sequenced
using Sanger dideoxy sequencing method. Of the 200 P. falciparum DNA samples
screened, sequencing for mutations in K13, pfmdr-2, fd, pfcrt, arps10 genes was successful
in 142, 186, 141, 128 and 74 samples respectively. Overall, we detected six (4.2%, 6/142;
95%CI: 1.4–7.0) K13 single nucleotide polymorphisms (SNPs), of which 3.9% (2/51), 4.4%
(4/91) occurred in 2005 and 2013 samples respectively. All four K13 SNPs in 2013 samples
were non-synonymous (A578S, E596V, S600C and E643K) while of the two SNPs in 2005
samples, one (Y588N) is non-synonymous and the other (I587I) is synonymous. There was
no statistically significant difference in the prevalence of K13 (p = 0.112) SNPs in the samples
collected in 2005 and 2013. The overall prevalence of SNPs in pfmdr-2 gene was
39.8% (74/186, 95%CI: 25.1–50.4). Of this, 4.2% (4/95), 76.9% (70/91) occurred in 2005
and 2013 samples respectively. In 2005 samples only one SNP, Y423F (4.2%, 4/95) was
found while in 2013, Y423F (38.5%, 35/91) and I492V (38.5%, 35/91) SNPs in the pfmdr-2
gene were found. There was a statistically significant difference in the prevalence of pfmdr-2
SNPs in the samples collected in 2005 and 2013 (p<0.001). The overall prevalence of
arps10 mutations was 2.7% (2/72, 95%CI: 0.3–4.2). Two mutations, V127M (4.5%: 1/22) and D128H (4.5%: 1/22) in the arps10 gene were each found in P. falciparum parasite samples
collected in 2013. There was no statistically significant difference in the prevalence of
arps10 SNPs in the samples collected in 2005 and 2013 (p = 0.238). There were more
pfmdr-2 SNPs in P. falciparum parasites collected after introduction of Artemisinin combination
therapies in malaria treatment. This is an indicator of the need for continuous surveillance
to monitor emergence of molecular markers of artemisinin resistance and its potential
drivers in malaria affected regions globally
Description
Keywords
Arps10, pfmdr-2, Gene mutations, Plasmodium falciparum
Citation
Ocan M, Ashaba FK, Mwesigwa S, Edgar K, Kamya MR, Nsobya SL (2022) Prevalence of arps10, fd, pfmdr-2, pfcrt and pfkelch13 gene mutations in Plasmodium falciparum parasite population in Uganda. PLoS ONE 17(5): e0268095. https://doi.org/10.1371/journal.pone.0268095