Molecular Method Optimization to Identify Plasmodium falciparum Multidrug Resistance 1 (pfmdr1) gene as a Predictor of Antimalarial Resistance

Erma Sulistyaningsih, Rosita Dewi, Sheilla Rachmania, Irawan Fajar Kusuma, Muhammad Rizqi Kholifaturrohmy, Yunita Armiyanti, Sakinatus Sariroh Kholifaturrohmah, Made Prasanti Andriani

Abstract


Several approaches have been designed to control malaria, a disease with high morbidity and mortality, but they face some hurdles. Antimalarial resistance is one of the major challenges for malaria elimination, so the detection of antimalarial resistance is essential. Several molecular markers for antimalarial resistance have been identified, including Plasmodium falciparum multidrug resistance 1 (pfmdr1) gene. This study determined the optimization of molecular techniques to identify the pfmdr1 gene as an antimalarial resistance predictor in Indonesia. The study included patients diagnosed with uncomplicated or severe malaria originating from the health district of Kerom Regency, Papua Province, and Dr. Soebandi Hospital, Jember, East Java Province. Blood samples were collected in the Whatmann filer paper after informed consent. DNA was isolated from dried blood filter paper, and nested PCR was performed using a specific primer, the pfmdr1-A and pfmdr1-B genes. The PCR cycle was optimized based on previous studies. The pfmdr1-A has a similar setting to the earlier study, but the pfmdr1-B had a different optimum setting from the previous study with the annealing temperature of 57oC for nested-1 and 62oC for nested-2. This PCR setting could be used for further examination. The positive results of the amplification indicated the potential for antimalarial resistance in the parasite population. A study on the gene copy number and polymorphism is essential to determine the definitive conclusion on antimalarial resistance.

Keywords


gene; malaria; pfmdr1, Plasmodium falciparum; resistance

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References


Adamu, A., Jada, M. S., Haruna, H. M. S., Yakubu, B. O., Ibrahim, M. A., Balogun, E. O., Sakura, T., Inaoka, D. K., Kita, K., Hirayama, K., Culleton, R., & Shuaibu, M. N. (2020). Plasmodium falciparum multidrug resistance gene-1 polymorphisms in Northern Nigeria: implications for the continued use of artemether-lumefantrine in the region. Malaria Journal, 19(1), 1–10. https://doi.org/10.1186/s12936-020-03506-z

Al-Mekhlafi, H. M., Madkhali, A. M., Abdulhaq, A. A., Atroosh, W. M., Ghzwani, A. H., Zain, K. A., Ghailan, K. Y., Hamali, H. A., Mobarki, A. A., Alharazi, T. H., Eisa, Z. M., & Lau, Y. L. (2022). Polymorphism analysis of pfmdr1 gene in Plasmodium falciparum isolates 11 years post-adoption of artemisinin-based combination therapy in Saudi Arabia. Scientific Reports, 12(1), 1–13. https://doi.org/10.1038/s41598-021-04450-x

Dondorp, A. M., Nosten, F., Yi, P., Das, D., Hanpithakpong, W., Ph, D., Lee, S. J., Ph, D., Ringwald, P., Ph, D., Imwong, M., Ph, D., Chotivanich, K., Ph, D., & Lim, P. (2009). Artemisinin Resistance in Plasmodium falciparum Malaria . New England Journal of Medicine, 361(17), 1714–1714. https://doi.org/10.1056/nejmx090050

Fairhurst, R. M. (2015). Understanding artemisinin-resistant malaria. Current Opinion in Infectious Diseases, 28(5), 417–425. https://doi.org/10.1097/qco.0000000000000199

Fitri, L. E., Pawestri, A. R., Winaris, N., Endharti, A. T., Khotimah, A. R. H., Abidah, H. Y., & Huwae, J. T. R. (2023). Antimalarial Drug Resistance: A Brief History of Its Spread in Indonesia. Drug Design, Development and Therapy, 17(April), 1995–2010. https://doi.org/10.2147/DDDT.S403672

Green, M. R., & Sambrook, J. (2019). Nested Polymerase Chain Reaction (PCR). Cold Spring Harbor Protocols, 2019(2). https://doi.org/10.1101/pdb.prot095182

Kemenkes, R. (2022). Rencana Kerja Tahunan Direktorat Jenderal Tenaga Kesehatan 2022. In Direktorat Jenderal Tenaga Kesehatan Kementrian Kesehatan Republik Indonesia (Vol. 1, Issue 2).

Manirakiza, G., Kassaza, K., Taremwa, I. M., Bazira, J., & Byarugaba, F. (2022). Molecular identification and antimalarial drug resistance profile of Plasmodium falciparum from patients attending Kisoro Hospital, southwestern Uganda. Malaria Journal, 21(1), 1–10. https://doi.org/10.1186/s12936-021-04023-3

Mungthin, M., Khositnithikul, R., Sitthichot, N., Suwandittakul, N., Wattanaveeradej, V., Ward, S. A., & Na-Bangchang, K. (2010). Association between the pfmdr1 gene and in Vitro artemether and lumefantrine sensitivity in thai isolates of Plasmodium falciparum. American Journal of Tropical Medicine and Hygiene, 83(5), 1005–1009. https://doi.org/10.4269/ajtmh.2010.10-0339

Noreen, N., Ullah, A., Salman, S. M., Mabkhot, Y., Alsayari, A., & Badshah, S. L. (2021). New insights into the spread of resistance to artemisinin and its analogues. Journal of Global Antimicrobial Resistance, 27, 142–149. https://doi.org/10.1016/j.jgar.2021.09.001

Nsanzabana, C., Djalle, D., Guérin, P. J., Ménard, D., & González, I. J. (2018). Tools for surveillance of antimalarial drug resistance: An assessment of the current landscape. Malaria Journal, 17(1), 1–16. https://doi.org/10.1186/s12936-018-2185-9

Price, R. N., Uhlemann, A. C., Brockman, A., McGready, R., Ashley, E., Phaipun, L., Patel, R., Laing, K., Looareesuwan, S., White, N. J., Nosten, F., & Krishna, S. (2004). Mefloquine resistance in Plasmodium falciparum and increased pfmdr1 gene copy number. Lancet, 364(9432), 438–447. https://doi.org/10.1016/S0140-6736(04)16767-6

QIAGEN. (2007). QIAamp ® DNA Mini and Blood Mini Handbook For DNA purification. 104(March), 101–108. http://www.ncbi.nlm.nih.gov/pubmed/16509153

Si, K., He, X., Chen, L., Zhang, A., Guo, C., & Li, M. (2023). The structure of Plasmodium falciparum multidrug resistance protein 1 reveals an N-terminal regulatory domain. Proc Natl Acad Sci U S A, 120(32), 2023. https://doi.org/10.1073/pnas

Sisowath, C., Ferreira, P. E., Bustamante, L. Y., Dahlström, S., Mårtensson, A., Björkman, A., Krishna, S., & Gil, J. P. (2007). The role of pfmdr1 in Plasmodium falciparum tolerance to artemether-lumefantrine in Africa. Tropical Medicine and International Health, 12(6), 736–742. https://doi.org/10.1111/j.1365-3156.2007.01843.x

Veiga, M. I., Dhingra, S. K., Henrich, P. P., Straimer, J., Gnädig, N., Uhlemann, A. C., Martin, R. E., Lehane, A. M., & Fidock, D. A. (2016). Globally prevalent PfMDR1 mutations modulate Plasmodium falciparum susceptibility to artemisinin-based combination therapies. Nature Communications, 7(May). https://doi.org/10.1038/ncomms11553

WHO. (2014). Severe Malaria. Trop Med Int Health, 19(Suppl I), 7–131.

WHO. (2023). World malaria report 2023. World Health Organization.

WHO, W. H. O. (2022). Strategy to respond to antimalarial drug resistance in Africa. In World Health Organization (Vol. 1).

WHO, W. H. O. (2024). WHO Guidelines for malaria. Who, November, 210.

Wurtz, N., Fall, B., Pascual, A., Fall, M., Baret, E., Camara, C., Nakoulima, A., Diatta, B., Fall, K. B., Mbaye, P. S., Diémé, Y., Bercion, R., Wade, B., & Pradines, B. (2014). Role of Pfmdr1 in In Vitro Plasmodium falciparum Susceptibility to Chloroquine, Quinine, Monodesethylamodiaquine, Mefloquine, Lumefantrine, and Dihydroartemisinin. Antimicrobial Agents and Chemotherapy, 58(12), 7032–7040. https://doi.org/10.1128/AAC.03494-14

Xu, C., Wei, Q., Yin, K., Sun, H., Li, J., Xiao, T., Kong, X., Wang, Y., Zhao, G., Zhu, S., Kou, J., Yan, G., & Huang, B. (2018). Surveillance of Antimalarial Resistance Pfcrt, Pfmdr1, and Pfkelch13 Polymorphisms in African Plasmodium falciparum imported to Shandong Province, China. Scientific Reports, 8(1), 1–9. https://doi.org/10.1038/s41598-018-31207-w




DOI: https://doi.org/10.14421/biomedich.2025.142.673-678

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Copyright (c) 2025 Erma Sulistyaningsih, Rosita Dewi, Sheilla Rachmania, Irawan Fajar Kusuma, Muhammad Rizqi Kholifaturrohmy, Yunita Armiyanti, Sakinatus Sariroh Kholifaturrohmah, Made Prasanti Andriani



Biology, Medicine, & Natural Product Chemistry
ISSN 2089-6514 (paper) - ISSN 2540-9328 (online)
Published by Sunan Kalijaga State Islamic University & Society for Indonesian Biodiversity.

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