PfCRT GENE POLYMORPHISMS IN PLASMODIUM FALCIPARUM ISOLATES FROM MALARIA PATIENTS IN MALARIA ENDEMIC AREAS USING ACT AS STANDARD THERAPY

Jhons Fatriyadi Suwandi; Betta Kurniawan; Hanna Mutiara; Aila Karyus

doi:10.32539/mks.v54i1.15137

PfCRT GENE POLYMORPHISMS IN PLASMODIUM FALCIPARUM ISOLATES FROM MALARIA PATIENTS IN MALARIA ENDEMIC AREAS USING ACT AS STANDARD THERAPY

Jhons Fatriyadi Suwandi, Betta Kurniawan, Hanna Mutiara, Aila Karyus

Abstract

The pfcrt gene is a biomarker to determine the resistance of Plasmodium falciparum to chloroquine and amodiaquine. When chloroquine was switched to ACT, it is very likely that there will be an increase in wild type strains (pfcrt K76) due to the absence of exposure to chloroquine. Currently chloroquine has not used for malaria treatment. The aims of this study were to identify polymorphisms in the pfcrt gene and phylogenetic analysis of Plasmodium falciparum isolates from malaria patients in Pesawaran Regency, Lampung Province. This research is a laboratory research by analyzing blood samples which are stored biological materials (BBT). DNA isolation was carried out using a DNA isolation kit QIAmp DNA Mini kit from Qiagen, followed by amplification using an appropriate primer. The amplification results were followed by sequencing and then analyzed using Mega 10. The results showed that all samples had mutations in codons 76 (K76T) and 72 (C72S) of the pfcrt gene. Pesawaran isolates were closely related to other isolates. The conclusion of this study is that polymorphisms were found in codons 72 and 76 of the PfCRT gene, although chloroquine has long been abandoned as an antimalarial. The sequenced pesawaran isolates were also related to other isolates.

Keywords

pfcrt, Plasmodium falciparum, chloroquinee, resistans

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References

Mayengue PI, Ndounga M, Malonga FV, Bitemo M, Ntoumi F. Genetic polymorphism of merozoite surface protein-1 and merozoite surface protein-2 in Plasmodium falciparum isolates from Brazzaville, Republic of Congo. Malar J. 2011;10(1):276.

doi:10.1186/1475-2875-10-276

World Health Organization. World Malaria Report. World Health Organization; 2020. doi:ISBN 978 92 4 1564403

Ministry of Health Republic of Indonesia. Indonesia Health Profile 2019. Ministry of Health Republic of Indonesia; 2020.

Health Office of Lampung Province. Lampung Province Health Profile 2019. Health Office of Lampung Province; 2020.

Ministry of Health Republic of Indonesia. Indonesia Health Profile 2008. Ministry of Health Republic of Indonesia; 2009.

Suwandi JF. Polymorphism of PfMDR1 and PfATP6 Genes in Plasmodium Isolates from Falciparum Malaria Patients in Pesawaran District. Published online 2014.

Syafruddin D, Asih PBS, Casey GJ, et al. Molecular epidemiology of Plasmodium falciparum resistance to antimalarial drugs in Indonesia. Am J Trop Med Hyg. 2005;72(2):174-181. http://www.ncbi.nlm.nih.gov/pubmed/15741554

Kamelia M. Study of The Plasmodium falciparum chloroquine resistance transporter (pfcrt) gene Polymorphism in Patients with Malaria in Bandar Lampung and South Lampung Regency, The Province of Lampung. Published online 2010.

Suwandi JF. Analysis of Plasmodium Falciparum PfCRT Gene Mutations in Malaria Patients in Lahat District Hospital. Faculty of Medicine University of Lampung; 2011.

Ministry of Health Republic of Indonesia. Guidelines for the Management of Malaria Cases in Indonesia. Ministry of Health Republic of Indonesia; 2008.

Gresty KJ, Gray KA, Bobogare A, et al. Genetic mutations in pfcrt and pfmdr1 at the time of artemisinin combination therapy introduction in South Pacific islands of Vanuatu and Solomon Islands. Malar J. 2014;13(1):1-9.

doi:10.1186/1475-2875-13-406

Sondo P, Derra K, Nakanabo SD, et al. Artesunate-amodiaquine and artemether-lumefantrine therapies and selection of pfcrt and PfmdR1 alleles in Nanoro, Burkina Faso. PLoS One. 2016;11(3). doi:10.1371/journal.pone.0151565

Bretscher MT, Dahal P, Griffin J, et al. The duration of chemoprophylaxis against malaria after treatment with artesunate-amodiaquine and artemether-lumefantrine and the effects of pfmdr1 86Y and pfcrt 76T: A meta-analysis of individual patient data. BMC Med. 2020;18(1):1-17. doi:10.1186/s12916-020-1494-3

Tajebe A, Aemero M, Francis K, Magoma G. Identification of chloroquine resistance Pfcrt-K76T and determination of Pfmdr1-N86Y copy number by SYBR Green I qPCR. Asian Pac J Trop Biomed. 2015;5(3):208-220. doi:10.1016/S2221-1691(15)30008-3

Ministry of Health Republic of Indonesia. Handbook of Malaria Case Management. Ministry of Health Republic of Indonesia; 2020. http://www.malaria.id/p/buku-malaria.html

Sambrook J, Fritsch EF, Maniatis. Molecular Cloning, A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory Press; 1989.

Humphreys GS, Merinopoulos I, Ahmed J, et al. Amodiaquine and Artemether-Lumefantrine Select Distinct Alleles of the Plasmodium falciparum mdr1 Gene in Tanzanian Children Treated for Uncomplicated Malaria ?. Antimicrob Agents Chemother. 2007;51(3):991-997. doi:10.1128/AAC.00875-06

Moll K, Ljungström I, Perlmann H, Scherf A, Wahlgren M. Methods in Malaria Research. 2008th ed. Malaria Research and Reference Reagent Resource Center, University Boulervard, Manassas; 2008.

Ibrahim ML, Khim N, Adam HH, Ariey F, Duchemin J-B. Polymorphism of PfATPase in Niger: detection of three new point mutations. Malar J. 2009;8:28. doi:10.1186/1475-2875-8-28

Mwai L, Ochong E, Abdirahman A, et al. Chloroquine resistance before and after its withdrawal in Kenya. Malar J. 2009;8(1):1-10.

doi:10.1186/1475-2875-8-106

Kublin JG, Cortese JF, Njunju EM, et al. Reemergence of chloroquine-sensitive Plasmodium falciparum malaria after cessation of chloroquine use in Malawi. J Infect Dis. 2003;187(12):1870-1875. doi:10.1086/375419

Atroosh WM, Al-Mekhlafi HM, Mahdy MAK, Surin J. The detection of pfcrt and pfmdr1 point mutations as molecular markers of chloroquine drug resistance, Pahang, Malaysia. Malar J. 2012;11(1):1.

doi:10.1186/1475-2875-11-251

DOI: https://doi.org/10.32539/mks.v54i1.15137