Genetic Insights into Plasmodium falciparum Resistance to Sulfadoxine-pyrimethamine through a Study of the 540-dihydropteroate Synthetase Gene Mutations in Jos, Nigeria

Nelson J. Nwankwo

Department of Biochemistry, University of Jos, Plateau State, Nigeria.

Joel Paul

Department of Biochemistry, University of Jos, Plateau State, Nigeria.

Shangshikmwa K. Gaknung

Department of Biochemistry, University of Jos, Plateau State, Nigeria.

Timloh Fakdul

Department of Biochemistry, University of Jos, Plateau State, Nigeria.

Mercy O. Simon

Department of Biochemistry, University of Jos, Plateau State, Nigeria.

Dinci T. Davou

Department of Biotechnology and Drug Development, NVRI, Plateau State, Nigeria.

Mary M. Mankilik

Department of Biochemistry, University of Jos, Plateau State, Nigeria.

Richard J. Kutshik

Department of Biochemistry, University of Jos, Plateau State, Nigeria.

Bitrus Yakubu

Department of Biotechnology and Drug Development, NVRI, Plateau State, Nigeria.

Ishaya Y. Longdet *

Department of Biochemistry, University of Jos, Plateau State, Nigeria.

*Author to whom correspondence should be addressed.


Aim: This study investigates mutations at codon 540 in the dihydropteroate synthetase (540dhps) gene that may confer resistance to sulfadoxine-pyrimethamine treatment by Plasmodium falciparum in Jos.

Study Design: The research employs a cross-sectional study design.

Place and Duration of Study: Samples were collected in Jos, Plateau State, Nigeria, from October 2019 to January 2021.

Methodology: One hundred malaria cases underwent assessment, and parasite DNA was extracted from whole blood samples using Zymo extraction kits. PCR and gel electrophoresis identified the Plasmodium genus and P. falciparum, along with the 540dhps gene. Sanger sequencing and bioinformatics analyses were performed.

Results: About 50% of the collected samples were positive for the Plasmodium genus, with P. falciparum in 96% of cases. About 16% of P. falciparum samples were successfully amplified for the 540dhps gene. A silent point mutation was discovered at position 10 of the nucleotide sequence of the 540dhps gene which did not result in amino acid changes.

Conclusion: The study highlights the predominant presence of Plasmodium falciparum in Jos, with a substantial 96% prevalence among malaria patients. Unlike the previously reported mutations at codon K540E on the 540dhps gene from other studies, the observed mutation on the nucleotide sequence did not affect the amino acid sequence. Thus, the observed mutation may not be associated with resistance to sulfadoxine-pyrimethamine drugs.

Keywords: Malaria, drug-resistance, single-nucleotide polymorphism, Plasmodium falciparum

How to Cite

Nwankwo , Nelson J., Joel Paul, Shangshikmwa K. Gaknung, Timloh Fakdul, Mercy O. Simon, Dinci T. Davou, Mary M. Mankilik, Richard J. Kutshik, Bitrus Yakubu, and Ishaya Y. Longdet. 2024. “Genetic Insights into Plasmodium Falciparum Resistance to Sulfadoxine-Pyrimethamine through a Study of the 540-Dihydropteroate Synthetase Gene Mutations in Jos, Nigeria”. Asian Journal of Biotechnology and Genetic Engineering 7 (1):128-38.


Download data is not yet available.


Tajebe A, Magoma G, Aemero M, & Kimani F. Detection of mixed infection level of Plasmodium falciparum and plasmodium vivax by SYBR Green I-based real-Time PCR in North Gondar, north-west Ethiopia. Malaria Journal. 2014;13(1),1–8.


UNICEF. Malaria data; 2021. Available:

Sondo P, Derra K, Lefevre T, Diallo-Nakanabo S, Tarnagda Z, Zampa O, Kazienga A, Valea I, Sorgho H, Ouedraogo JB, Guiguemde TR, Tinto H. Genetically diverse Plasmodium falciparum infections, within-host competition and symptomatic malaria in humans. Scientific Reports. 2019;9(1):1–9. Available:

Joste V, Maurice L, Bertin G I, Aubouy A, Boumédiène F, Houzé S, Ajzenberg D, Argy N, Massougbodji A, Dossou-Dagba I, Alao MJ, Cot M, Deloron P, Faucher JF. Identification of Plasmodium falciparum and host factors associated with cerebral malaria: Description of the protocol for a prospective, case-control study in Benin (NeuroCM). BMJ Open. 2019;9(5),1–9.


Moyeh MN, Njimoh DL, Evehe MS, Ali IM, Nji AM, Nkafu DN, Masumbe PN, Barbara A, Ndikum VN, Mbacham WF. Effects of drug policy changes on evolution of molecular markers of Plasmodium falciparum resistance to chloroquine, Amodiaquine, and sulfadoxine-pyrimethamine in the south west region of cameroon. Malar Res Treat. 2018; 2:7071383. DOI: 10.1155/2018/7071383

Baraka V, Ishengoma DS, Fransis F, Minja DTR, Madebe RA, Ngatunga D, Geertruyden JPV. High level Plasmodium sulfadoxine - pyrimethamine resistance with the concomitant occurrence of septuple haplotype in Tanzania. Malaria Journal. 2015;14:439. Available:

Menegon M, Pearce RJ, Inojosa WO, Pisani V, Abel PM, Matondo A, Bisoffi Z, Majori G, Ord R, Warhurst DC, Roper C, & Severini C. Monitoring for multidrug-resistant Plasmodium falciparum isolates and analysis of pyrimethamine resistance evolution in Uige province, Angola. Tropical Medicine & International Health. 2009;14(10):1251–1257.


Mwanza S, Joshi S, Nambozi M, Chileshe J, Malunga P, Kabuya JBB, Hachizovu S, Manyando, C, Mulenga M, Laufer M. The return of chloroquine-susceptible Plasmodium falciparum malaria in Zambia. Malaria Journal. 2016;15(584),2–6. Available:

Kavishe RA, Kaaya RD, Nag S, Krogsgaard C, Notland JG. Molecular monitoring of Plasmodium falciparum super resistance to sulfadoxine – pyrimethamine in Tanzania. Malaria Journal. 2016;15:335. Available:

Marks F, Evans J, Meyer CG, Browne EN, Flessner C, von Kalckreuth V, Eggelte TA, Horstmann RD, May J. High prevalence of markers for sulfadoxine and pyrimethamine resistance in Plasmodium falciparum in the absence of drug pressure in the Ashanti region of Ghana. Antimicrob Agents Chemother. 2005;49(3):1101-1105. DOI: 10.1128/AAC.49.3.1101-1105.2005

Lumb V, Das MK, Singh N, Dev V, Khan W, & Sharma Y D. Multiple origins of Plasmodium falciparum Dihydropteroate synthetase mutant alleles associated with sulfadoxine resistance in India. Antimicrob Agents Chemother. 2011;55(6),2813–2817.


Awono-ambe PH. Prevalence of Plasmodium falciparum parasites resistant to sulfadoxine/pyrimethamine in pregnant women in Yaounde Cameroon: emergence of highly resistant pfdhfr / 540dhps alleles.2015;2566–2571. Available:

Mayxay M, Pukrittayakamee S, Newton PN, White NJ. Mixed-species malaria infections in humans. Trends in Parasitology. 2004; 20(5). Available:

Bloland & WHO. Anti-Infective drug resistance surveillance and containment team: Drug resistance in malaria. A background document for the WHO global strategy for containment of antimicrobisl resistance. 2001;4:27,

Available: Accessed November 18, 2019.

Iwalokun BA, Iwalokun SO, Adebodun V, Balogun M. Carriage of mutant dihydrofolate reductase and dihydropteroate synthase genes among Plasmodium falciparum Isolates recovered from pregnant women with asymptomatic infection in Lagos, Nigeria. Med Princ Pract. 2015;24:436-443.

DOI: 10.1159/000430987

Plebani Mario. Laboratory-associated and diagnostic errors: A neglected link Diagnosis. 2014;1(1):89-94.


Jiang T, Chen J, Fu H. et al. High prevalence of Pfdhfr–540dhps quadruple mutations associated with sulfadoxine–pyrimethamine resistance in Plasmodium falciparum isolates from Bioko Island, Equatorial Guinea. Malar J. 2019;18,101.


Saba S, Akhtar T, Hanif A, Sahar S, Niaz S, Bilal H. Molecular detection of malaria in south punjab with higher proportion of mixed infections. Iranian Journal of Parasitology. 2014;9(1),37–43.

Paul J, Nwankwo NJ, Simon M, et al. Non-synonymous mutations associated with Plasmodium falciparum artemisinin resistant gene (Pfkelch13) in Malaria cases, Jos Nigeria. Journal of Advances in Biology & Biotechnology. 2023;26(10): 28-38.


Oriero EC, Olukosi AY, Oduwole OA, Djimde A, D'Alessandro U, Meremikwu MM, Amambua-Ngwa A. Seroprevalence and parasite rates of plasmodium malariae in a High malaria transmission setting of Southern, Nigeria. The American Journal of Tropical Medicine and Hygiene, 2020;103(6),2208–2216.


Oboh MA, Badiane AS, Ntadom G, et al. Molecular identification of plasmodium species responsible for malaria reveals plasmodium vivax isolates in duffy negative individuals from southwestern Nigeria. Malar J. 2018;17,439. Available:

Muller O. Challenges for control and elimination in the 21st century. Malaria Afri 2011;60:193.

Muller O. Challenges for control and elimination in the 21st century. Malaria Afri 2011;60:193.

Shibeshi MA, Kifle ZD, Atnafie SA. Antimalarial drug resistance and novel targets for antimalarial drug discovery. Infect Drug Resist. 2020;13:4047-4060. Available:

Alam MT, De Souza DK, Vinayak S, et al. Selective sweeps and genetic lineages of Plasmodium falciparum drug-resistant alleles in Ghana. J Infect Dis. 2011; 203:220–227.

Dickson ET, Hyman P Mutation, Silent. Brenner's Encyclopedia of Genetics. 2013;553-555. Available:

Doumbo S, Ongoiba OA, Doumtabé D, et al. Prevalence of Plasmodium falciparum , anemia and molecular markers of chloroquine and sulfadoxine-pyriméthamine resistance in delivered women in Fana, Mali (in French). Bull Soc Pathol Exot. 2013;106:188–192.

Gatton ML, Martin LB, Cheng Q. Evolution of resistance to sulfadoxine-pyrimethamine in Plasmodium falciparum. PMC Journal. 2004;48(6),2116–2123. Available:

Gosling RD, Cairns ME, Chico RM, Chandramohan D. Intermittent preventive treatment against malaria: an update, Expert Review of Anti-infective Therapy. 2010;8:5,589-606,