Polymorphisms in the pfcrt and pfmdr1 genes in Plasmodium falciparum isolates from South Sumatera, Indonesia

Irsan Saleh, Dwi Handayani, Chairil Anwar



DOI: https://doi.org/10.13181/mji.v23i1.679

Abstract


Background: Over the past decade, antimalarial drug resistance has rapidly become a major public health problem in South East Asia region including South Sumatra. This study aimed to determine the extent of gene polymorphisms associated with chloroquine resistance (CQR) in P. falciparum isolates from Lahat, Sekayu, Baturaja and Palembang district.

Methods: A molecular study was conducted to identify the mutant alleles of the genes associated with the resistance to chloroquine among the isolates of Plasmodium falciparum from South Sumatera. Blood from 25 patients was collected, DNA was isolated, and the sequences of two different genes (Plasmodium falciparum chloroquine resistance transporter/pfcrt and Plasmodium falciparum multidrug resistance/pfmdr1) were analyzed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP).

Results: This study identified polymorphism in the pfcrt 76-Thr in all isolates and pfmdr1 86-Tyr. These findings may reflect the failure of treatment with the standard dose of chloroquine within the last few years in South Sumatera.

Conclusion: PCR-RFLP technique provide a simple and rapid method of detecting polymorphisms in genes that may predict chloroquine resistance (CQR). Although the identification of the polymorphism in the pfcrt and pfmdr1 genes provides a significant indicator of CQR, further studies are needed to determine the role of these polymorphisms in the in vivo and in vitro responses to drug treatment.

Keywords: chloroquine, Plasmodium falciparum, pfmdr1, pfcrt


Full Text:

PDF

References


  1. World Health Organization. Guidelines for the treatment of malaria. Geneva: World Health Organization; 2006.
  2. World Health Organization. World Malaria Report. Geneva: World Health Organization; 2010.
  3. Dinas Kesehatan Provinsi Sumatera Selatan. Pencegahan penyakit dan penyehatan lingkungan. Profil Kesehatan Provinsi Sumatera Selatan. 2010. Indonesian.
  4. Fairhurst RM, Nayyar GML, Breman JG, Hallet R, Vennerstorm JL, Duong S, et al. Artemisinine-resistant malaria: research challenges, opportunities, and public health implications. Am J Trop Med Hyg. 2012;87(2):231-41. http://dx.doi.org/10.4269/ajtmh.2012.12-0025
  5. Ebisawa I, Fukuyama T. Chloroquine-resistant of Plasmodium falciparum malaria from West Irian and East Kalimantan. Ann Trop Med Parasitol. 1975;69(3):275-9.
  6. Syafruddin D, Asih PB, Casey GJ, Maguire J, Baird JK, Nagesha HS, et al. Molecular epidemiology of Plasmodium falciparum resistance to antimalarial drugs in Indonesia. Am J Trop Med Hyg. 2005;72(2):174-81.
  7. Sutar SK, Gupta B, Ranjit M, Kar SK, Das A. Sequence analysis of coding DNA fragments of pfcrt and pfmdr-1 genes in Plasmodium falciparum isolates from Odisha, India. Mem Inst Oswaldo Cruz. 2011;106(1): 78-84. http://dx.doi.org/10.1590/S0074-02762011000100013
  8. Wellems TE. Molecular genetics of drug resistance in Plasmodium falciparum malaria. Parasitol Today. 1991;7(5):110-2. http://dx.doi.org/10.1016/0169-4758(91)90168-N
  9. Carlton J, Fidock DA, Djimde A, Plowe CV, Baker J, Peters JM, et al. Conservation of a novel vacuolar transporter in Plasmodium species and its role in chloroquine resistance in falciparum but not vivax malaria. Curr Opin Microbiol. 2001;4(4):415-20. http://dx.doi.org/10.1016/S1369-5274(00)00228-9
  10. Syafruddin D, Asih PBS, Aggarwal SL, Shankar A. Frequency distribution of antimalarial drug-resistant alleles among isolates of Plasmodium falciparum in Purworejo district, Central Java Province, Indonesia. Am J Trop Med Hyg. 2003;69(6):614-20.
  11. Wooden J, Kyes S, Sibley CH. PCR and strain identification in Plasmodium falciparum. Parasitol Today. 1993;9(8):303-5. http://dx.doi.org/10.1016/0169-4758(93)90131-X
  12. Duraisingh MT, Drakeley CJ, Muller O, Bailey R, Snounou G, Targett GA, et al. Evidence for selection for the tyrosine-86 allele of the pfmdr1 gene of Plasmodium falciparum by chloroquine and amodiaquine. Parasitology. 1997;114:205-11. http://dx.doi.org/10.1017/S0031182096008487
  13. Scopel KK, Fontes CJ, Nunes AC, Horta MF, Braga EM. Low sensitivity of nested PCR using Plasmodium DNA extracted from stained thick blood smears: an epidemiological retrospective study among subjects with low parasitaemia in an endemic area of the Brazilian Amazon region. Malar J. 2004;3(8):1-6.
  14. Wellems TE, Plowe CV. Chloroquine-resistant malaria. J Infect Dis. 2001;184(6):770-6. http://dx.doi.org/10.1086/322858
  15. Fidock DA, Nomura T, Talley AK, Cooper RA, Dzekunov SM, Ferdig MT, et al. Mutations in the P. falciparum digestive vacuole transmembrane protein PfCRT and evidence for their role in chloroquine resistance. Moll Cell. 2000;6(4):861-71. http://dx.doi.org/10.1016/S1097-2765(05)00077-8
  16. Chaijaroenkul W, Ward SA, Mungthin M, Johnson D, Owen A, Bray PG, et al. Sequence and gene expression of chloroquine resistance transporter (pfcrt) in the association of in vitro drugs resistance of Plasmodium falciparum. Malaria J. 2011; 10(42):1-9.
  17. Djimdé A, Doumbo OK, Cortese JF, Kayentao K, Doumbo S, Diourté Y, et al. A molecular marker for chloroquine resistant falciparum malaria. N Engl J Med. 2001;344(4):257-63. http://dx.doi.org/10.1056/NEJM200101253440403
  18. Dorsey G, Kamya MR, Singh A, Rosenthal PJ. Polymorphisms in the Plasmodium falciparum pfcrt and pfmdr-1 genes and clinical response to chloroquine in Kampala, Uganda. J Infect Dis. 2001;183(9):1417-20. http://dx.doi.org/10.1086/319865
  19. Thomas SM, Ndir O, Dieng T, Mboup S, Wypij D, Maguire JH, et al. In vitro chloroquine susceptibility and PCR analysis of pfcrt and pfmdr1 polymorphisms in Plasmodium falciparum isolates from Senegal. Am J Trop Med Hyg. 2002;66(5):474-80.
  20. Foote SJ, Thompson JK, Cowman AF, Kemp DJ. Amplification of multidrug resistance gene in some chloroquine resistant isolates of Plasmodium falciparum. Cell. 1989; 57(6):921-30. http://dx.doi.org/10.1016/0092-8674(89)90330-9
  21. Foote SJ, Kyle DE, Martin RK, Oduola AMJ, Forsyth K, Kemp DJ, et al. Several alleles of the multidrug-resistance gene are closely linked to chloroquine resistance in Plasmodium falciparum. Nature. 1990; 345:255-8. http://dx.doi.org/10.1038/345255a0
  22. Basco LK, Ndounga M, Ngane VF, Soula G. Molecular epidemiology of malaria in Cameroon. XIV. Plasmodium falciparum chloroquine resistance transporter (PFCRT) gene sequences of isolates before and after chloroquine treatment. Am J Trop Med Hyg. 2002;67(4): 392-5.
  23. Nagesha HS, Syafruddin D, Casey GJ, Susanti AI, Fryauff DJ, Reeder JC, et al. Mutations in the pfmdr1, dhfr and dhps genes of Plasmodium falciparum are associated with in-vivo drug resistance in West Papua, Indonesia. Trans R Soc Trop Med Hyg. 2001;95(1):43-9. http://dx.doi.org/10.1016/S0035-9203(01)90329-3
  24. Sutanto I, Supriyanto S, Ruckert P, Purnomo, Maguire JD, Bangs MJ. Comparative efficacy of chloroquine and sulfadoxine-pyrimethamine for uncomplicated Plasmodium falciparum malaria and impact on gametocyte carriage rates in the East Nusatenggara province of Indonesia. Am J Trop Med Hyg. 2004;70(5):467-73.





Copyright (c) 2014 Irsan Saleh, Dwi Handayani, Chairil Anwar

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

All articles and issues in Medical Journal of Indonesia have unique DOI number registered in Crossref.
 
Romeo