KnE Life Sciences
ISSN: 2413-0877
The latest conference proceedings on life sciences, medicine and pharmacology.
Experimental Models Point Mutations In Plasmodium falciparum pfatpase6 Gene Exposed to Recuring Artemisinin In Vitro
Published date: Dec 03 2017
Journal Title: KnE Life Sciences
Issue title: The Veterinary Medicine International Conference (VMIC)
Pages: 422-435
Authors:
Abstract:
The aims of this research to prove that repeated exposure of artemisinin can cause pfatpase6 gene mutation on Plasmodium falciparum in vitro. The research methods used culture In Vitro Plasmodium falciparum of strain 2300 IC50 value determination test artemisinin, artemisinin repeated exposure test (PO1, PO2, PO3 dan PO4) dose IC50, DNA extraction, gene amplification of pfatpase6 using Polymerase Chain Reaction (PCR) technique, electrophoresis, PCR product purification, labeling DNA from PCR results, DNA precipitation of PCR product, application of product labeling on the sequencing machines, analysis of the results of sequencing, and Data Analysis. The results of PCR pfatpase6 gene amplification include region 6 – 3216 for codon 89-1031 located in exon 1 and 2 Plasmodium falciparum 2300 by using five pairs of primers. Primer pair 1FR produce a long amplicon of 737 bp which covers of codon 89; primer pair 2FR produce a long amplicon of 813 bp which covers of codon 263, 431; primer pair 4FR produce a long amplicon of 700 bp which covers of codon 460, 465, 623; primer pair 5FR produce a long amplicon of 550 bp which includes of codon 683, 769; and primer pair 6FR produce a long amplicon of 876 bp which covers of codon 898, 1031.Multialigment pfatpase6 gene Plasmodium falciparum of strains Papua 2300 point mutations are obtained in the form of transition and transversion in treatment groups at the same nucleotide region 123, 2035, 2043, 2138 dan 2148. Conclusion of this research Artemisinin repeated exposure can cause point mutations in pfatpase6 genes Plasmodium falciparum of strains 2300 in vitro
Keyword: Artemisinin, Plasmodium falciparumof strain Papua 2300, pfatpase6 gene, point mutation
References:
1. World Health Organization. 2010. Global report on antimalarial drug efficacy and drug resistance 2000-2010. Geneva. 32
2. Veiga MI, Ferreira PE, Schmidt BA, Schmidt BA, Ribacke U, Bjorkman A, Tichopad A, Gil JP. Antimalarial exposure delays Plasmodium falciparum intra erytrocytic cycle and drives drug transporter Genes expression. Plos One .2010; 5 (8) : e12408.
3. Ullastres A, Merenciano M, Guro IU, Gonzales J. 2016. Stress and envirronmental regulation of gene expression and adaptation in bacteria : Transposable elements : a toolkit for stress and environmental adaptation in bacteria. John Willey & Sons Inc .Newjwersy . Vol 1 . Chapter 2.10.137-140.
4. Bla B.K, Yavo W, Trebissou J, Kipre R.G, Yapi F.H, N’guessan J.N. Djaman J.A. 2014. Polymorphisms of the Pfatpase6 and Pfcrt gene and their relationship with the in vitro susceptibility to dihydroartemisinin and chloroquine of Plasmodiumfalciparum isolates from Abobo, Côte d’Ivoire. Annals Parasitology .60(40) :259-266.
5. Globler L, Chavchich M, Haynes R.K, Edstein M.D,Grobler A .2014.Assessment of the induction of dormant ring stages in Plasmodium falciparum parasites by artemisone and artemisone entrapped in pheroid vesicles in vitro. Antimicrobial Agents and Chemotherapy. 58 (12): 7579-7582
6. Isozumi R, Uemura H, Kimata I, Ichinose Y, Logedi J, Omar A.H, Kaneko A. 2015. Novel mutations in K13 propeller gene of artemisinin-resistant Plasmodium falciparum.Emerging Infectious Diseases . 21 (3) :
7. Mugittu K, Genton B, Mshinda H and Beck HP. Molecular monitoring of Plasmodium falciparum resistance to artemisinin in Tanzania. Malaria Journal. 2006; 5:126
8. Jambou R, Legrand E, Niang M, Khim N, Lim P, Volney B, Ekala MT, Bouchier C, Esterre P, Fandeur T, Mercereau Pujijalon O. Resistance of P.falciparum field isolates to In vitro artemether and point mutations of the SERCA type Pf ATPase6. The Lancet. 2005; 366:1960-1963
9. Price RN, Uhlemann AC, Brockman A, McGready R, Ashley E,Phaipun L, Patel R, Laing K, Looareesuwan S, White NJ, Nosten F, Krishna S.Mefloquine resistance in Plasmodium falciparum and increased pfmdr1gene copy number. The Lancet. 2004; 364 : 438-447
10. Ferreira ID, Lopes D, Martenelli A, Ferreira C, Rosario VE, Cravo P.In vitro assessment of artesunate, artemether and amodiaquine susceptibility and molecular analysis of putative resistance associated mutation of Plasmodium falciparum from Sao Tome and Principe. Tropical Medicine and International. 2007; 12: 353-362
11. Dahlstrom S, Veiga MI, Ferreira P, Martensson A, Kaneko A, Anderson B, Bjorkman A, Gil JP. Diversity of the sarco/endoplasmic reticulum Ca(2+) ATPase6 orthologue of Plasmodium falciparum (PfATP6).Infect Genet6 Evol . 2008; 8:340-345
12. Zakary S, Hemati S2, Pirahmadi S, Afsharpad M, Raeisi A and Djadid ND. Molecular assessment of atpase6 mutations associated with artemisinin resistance among unexposed and exposed Plasmodium falciparum clinical isolates to artemisinin-based combination therapy Malaria Journal 2012, 11:37
13. Wilairat P, Kumpornsin K, Chookajorn T. 2016. Plasmodium falciparum malaria: Convergent evolutionary trajectories towards delayed clearance following artemisinin treatment. Medical Hypotheses 90:19–22
14. Paloque L, Ramadani A.P, Mercereau‑Puijalon O, Augereau J.M, Benoit‑Vical1 F.2016. Plasmodium falciparum: multi faceted resistance to artemisinins.Malar J 15:149
15. Njokah M.J, Kang J.N, Kinyua J, Kariuki D, Kimani F.T. 2016.In vitro selection of Plasmodiumfalciparum Pfcrt and Pfmdr1 variantsby artemisinin.Malar J . 15:381.
16. Trager W and Jensen J.B. Human malaria parasites in continuous culture. Science 1976; 193: 673-675
17. Yuan L, Hao M, Wu L, Zhao Z, Rosental B.M, Li X, He Y, Sun L, Feng G, Xiang Z, Cui L and Yang Z. 2014 Refrigeration provides a simple means to synchronize in vitrocultures of Plasmodium falciparum.Exp Parasitol. 140: 18–23. doi:10.1016/j.exppara.2014.03.010.
18. Garcia, L. Determination of parasitemia : Diagnostic medical parasitology 2007; ed. 5. ASM Press.Washington DC.
19. Ljungstrom I, Perlmann H, Schlictherle M, Scherf A, Wahlgren M. Methods in Malaria Research .2004; Ed 4. 1-240
20. Sanz LM, Crespo B, De-cozar C, Ding XC, Liergo,JL, Burrows JN, Garcia –Butos JF, Gamo FJ.P.falciparumin vitro killing rates allow to discriminate between different antimalarial mode of action. Plos One. 2012; 7(2): e30949
21. Imwong M, Dondorpn AM, Nosten F, Yi P, et al. Exploring the contribution of candidate genes to artemisinin resistance in P.falciparum. Antimicrobial Agents and Chemotherapy. 2010; 54 (7) : 2886-2892
22. Maslachah L, Dachlan Y.P, Nidom C.A, Fitri L.E. 2015. Phenotypic Profile of Plasmodium falciparum Papua 2300 strain Exposed to in vitro Antimalaria Artemisinin . MKB. 47(1).1-9.
23. Afoakwah R, Boampong JN, Acheampong DO, and Nwaefuna EK.Polymorphisms in Plasmodium falciparum adenosine triphosphatase6 (pfatpase6) gene and their significance in finding the genetic marker for artemisinin resistance.European Journal of Experimental Biology. 2011; 1(3): 7-13
24. Ibrahim ML, Khim N, Adam HH, Ariey F, Duchemin JB. Polymorphism of pfatpase 6 in Niger: detection of three new point mutation. Malaria Journal.2008; 8(28): 1-4
25. Miao M, Wang Z, Yang Z, Yuan L, Parker D.M, Putaporntip C, Jongwutiwes S, Xangsayarath P, Pongvongsa T, et al. . Genetic diversity and lack of artemisinin selection signature on the Plasmodium falciparum ATP in greater Mekong Subregion. Plos One.2013; 8 (3):e59192
26. Valderramos A..G, Fidock D..A. Transporters involved in resistance to antimalarial drugs. Trend in Pharmacological Science. 2006; 27(11): 594-601
27. Tahar R, Ringwald P, Basco LK. Molecular Epidemiology of malaria in Camerroon XXVIII in vitro activity of dihidroartemisinin against clinical isolates of Plasmodium falciparum and sequence analysis of the P.falciparumATPase6 Gene. Am.J.Trop Med.Hyg.2009; 81 (1): 13-18
28. Bertaux L, Quang LH, Sinou V, Thanh NX, Parzy D. New PfATPase6 mutation found in Plasmodium falciparum isolates from Vietnam. Antimicrobial Agent and Chemotherapy.2009; 53 (10) : 4570-4571
