Characterization of putative drug resistant biomarkers in Plasmodium falciparum isolated from Ghanaian blood donors

Abstract

Background: Plasmodium falciparum parasites, which could harbour anti-malaria drug resistance genes, are commonly detected in blood donors in malaria-endemic areas. Notwithstanding, anti-malaria drug resistant biomarkers have not been characterized in blood donors with asymptomatic P. falciparum infection. Methods: A total of 771 blood donors were selected from five districts in the Greater Accra Region, Ghana. Each donor sample was screened with malaria rapid diagnostic test (RDT) kit and parasitaemia quantified microscopically. Dried blood spots from malaria positive samples were genotyped for P. falciparum chloroquine resistance transporter (Pfcrt), P. falciparum multi-drug resistance (Pfmdr1), P. falciparum dihydropteroate-synthetase (Pfdhps), P. falciparum dihydrofolate-reductase (Pfdhfr) and Kelch 13 propeller domain on chromosome 13 (Kelch 13) genes. Results: Of the 771 blood donors, 91 (11.8%) were positive by RDT. Analysis of sequence reads indicated successful genotyping of Pfcrt, Pfmdr1, Pfdhfr, Pfdhps and Kelch 13 genes in 84.6, 81.3, 86.8, 86.9 and 92.3% of the isolates respectively. Overall, 21 different mutant haplotypes were identified in 69 isolates (75.8%). In Pfcrt, CVIET haplotype was observed in 11.6% samples while in Pfmdr1, triple mutation (resulting in YFN haplotype) was detected in 8.1% of isolates. In Pfdhfr gene, triple mutation resulting in IRNI haplotype and in Pfdhps gene, quintuple mutation resulting in AGESS haplotype was identified in 17.7% parasite isolates. Finally, five non-synonymous Kelch 13 alleles were detected; C580Y (3.6%), P615L (4.8%), A578S (4.8%), I543V (2.4%) and A676S (1.2%) were detected. Conclusion: Results obtained in this study indicated various frequencies of mutant alleles in Pfcrt, Pfmdr1, Pfdhfr, Pfdhps and Kelch 13 genes from P. falciparum infected blood donors. These alleles could reduce the efficacy of standard malaria treatment in transfusion-transmitted malaria cases. Incorporating malaria screening into donor screening protocol to defer infected donors is therefore recommended.

A review of the frequencies of Plasmodium falciparum Kelch 13 artemisinin resistance mutations in Africa

“…1 ) ( Page et al, 2021 ) undertaken in Africa from samples obtained from 2012 to 2020, across 30 countries ( Table S1 , Full table accessible at Harvard Dataverse: https://doi.org/10.7910/DVN/EUPXCF ). The Pfk13 genotypes detected across many African countries were primarily wild-type with only Ghana reporting the C580Y mutation ( Matrevi et al, 2019 ; Aninagyei et al, 2020 ) associated with the majority of ART resistance in SE Asia. There were no Pfk13 mutations reported in Benin (2014) , The Gambia (2012–2014) and Liberia (2014–2017) ( Table S1 ).…”

The newly discovered role of endocytosis in artemisinin resistance

“…30,[35][36][37] Additionally, K13 resistance mutations, including C580Y, which is the most prominent mutation in Southeast Asia, were detected in Cameroon 38 and Ghana. 39 Despite these reports of potentially resistant parasites in Africa, significant clinical consequences such as the clearance delay on par with those observed in Southeast Asia have not been reported in African settings.…”

“…35 In fact, K13 resistance mutations were initially found to be absent in Africa, but have now been detected in several African countries. 36,38,39,[139][140][141][142] One hypothesis to explain the occurrence of ART resistance mainly in Southeast Asia proposed a specific genetic background is required for the K13 mutation to arise. 143 However, in vitro experiments showed that K13 mutations can be introduced in parasites of African origin and confer in vitro resistance.…”

The emergence of chloroquine-sensitive Plasmodium falciparum is influenced by selected communities in some parts of the Central Region of Ghana

“…Hence n = z 2 pq/d2 n = (1.962) (0.884) (0.116)/(0.052), n = 0.39393/0.0025, n = 157.57. However, 214 P. falciparum parasites were analysed from the four study sites [ 19 , 20 ].…”