malaria; pfcrt; drug resistance; RT-PCR; sequencing; DR Congo
Abstract :
[en] BACKGROUND: In 2001, the World Health Organization (WHO) has recommended the use of artemisinin-based combination therapy (ACT) as the first-line treatment of uncomplicated malaria cases, as monotherapies had become ineffective in many parts of the world. As a result, the Democratic Republic of Congo (DRC) withdrew chloroquine (CQ) from its malaria treatment policy in 2002 and an artesunate (AS)-amodiaquine (AQ) combination became the ACT of choice in DRC in 2005. AQ-resistance (AQR) has been reported in several parts of the world and mutations in codons 72-76 of the Plasmodium falciparum chloroquine-resistance transporter (pfcrt) gene have been strongly correlated with resistance, especially mutations encoding the SVMNT haplotype. This haplotype was first identified in Southeast Asia and South America but was recently reported in two African countries neighbouring DRC. These facts raised two questions: the first about the evolution of CQ resistance (CQR) in DRC and the second about the presence of the SVMNT haplotype, which would compromise the use of AQ as a partner drug for ACT.
METHODS: A total of 213 thick blood films were randomly collected in 2010 from a paediatric clinic in Kinshasa, DRC. Microscopy controls and real-time polymerase chain reaction (RT-PCR) were performed for Plasmodium species identification. Haplotypes of the pfcrt gene were determined by sequencing.
RESULTS: The K76T mutation was detected in 145 out of 198 P. falciparum-positive samples (73.2%).In these 145 resistant strains, only the CVIET haplotype was detected.
CONCLUSIONS: This study is the first to assess the molecular markers of resistance to CQ and AQ after the introduction of ACT in DRC. The results suggest first that CQR is decreasing, as wild-type pfcrt haplotypes were found in only 26.8% of the samples and secondly that the SVMNT haplotype is not yet present in Kinshasa, suggesting that AQ remains valid as a partner drug for ACT in this region.
Disciplines :
Immunology & infectious disease
Author, co-author :
Mvumbi, Dieudonné; University of Kinshasa, DR Congo > Basic Sciences, School of Medicine > 1Biochemistry and Molecular Biology Unit, > 3
BOREUX, Raphaël ; Centre Hospitalier Universitaire de Liège - CHU > Microbiologie médicale
SACHELI, Rosalie ; Centre Hospitalier Universitaire de Liège - CHU > Microbiologie médicale
Mvumbi, Lelo; University of Kinshasa, DR Congo > Department of Basic Sciences, School of Medicine, > 1Biochemistry and Molecular Biology Unit,
Bobanga, Lengu; University of Kinshasa, DR Congo > Department of Tropical Medicine and Infectious Disease, School of Medicine
Situakibanza, Hippolyte; Department of Tropical Medicine and Infectious Disease, School of Medicine, University of Kinshasa, DR Congo > Department of Tropical Medicine and Infectious Disease, School of Medicine
Melin, Pierrette ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Bactériologie, mycologie, parasitologie, virologie
Kayembe, Jean-Marie; University of Kinshasa, DR Congo > Department of Internal Medicine, School of Medicine
Kalala, Lunganza; University of Kinshasa, DR Congo > 1Biochemistry and Molecular Biology Unit, Department of Basic Sciences, School of Medicine
De Mol, Patrick ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Département des sciences biomédicales et précliniques
Hayette, Marie-Pierre ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Bactériologie, mycologie, parasitologie, virologie
Language :
English
Title :
Assessment of pfcrt 72-76 haplotypes eight years after chloroquine withdrawal in Kinshasa, Democratic Republic of Congo
Alternative titles :
[fr] Détermination des haplotypes pfcrt 72-76, 8 ans après abandon de l'utilisation de la chloroquine à Kinshasa, RDC
Publication date :
20 December 2013
Journal title :
Malaria Journal
eISSN :
1475-2875
Publisher :
BioMed Central, United Kingdom
Volume :
12
Pages :
459
Peer reviewed :
Peer Reviewed verified by ORBi
Name of the research project :
Epidémiologie des résistances de Plasmodium falciparum en RDC
Funders :
CUD - Commission Universitaire pour le Développement
World Health Organization, Antimalarial drug combination therapy. Report of a WHO technical consultation Geneva: WHO 2001
Plasmodium chloroquine resistance and the search for a replacement antimalarial drug. Wellems TE, Science 2002 298 124 126 10.1126/science.1078167 12364789
Mutations in the P. falciparum digestive vacuole transmembrane protein PfCRT and evidence for their role in chloroquine resistance. Fidock DA, Nomura T, Talley AK, Cooper RA, Dzekunov SM, Ferdig MT, Ursos LM, Sidhu AB, Naude B, Deitsch KW, Su XZ, Wootton JC, Roepe PD, Wellems TE, Mol Cell 2000 6 861 871 10.1016/S1097-2765(05)00077-8 11090624
Chloroquine resistance in Plasmodium falciparum malaria parasites conferred by pfcrt mutations. Sidhu AB, Verdier-Pinard D, Fidock DA, Science 2002 298 210 213 10.1126/science.1074045 12364805
A molecular marker for chloroquine resistant falciparum malaria. Djimde A, Doumbo OK, Cortese JF, Kayentao K, Doumbo S, Diourte Y, Dicko A, Su X-Z, Noruma T, Fidock DA, Wellems TE, Plowe CV, N Engl J Med 2001 344 257 263 10.1056/NEJM200101253440403 11172152 (Pubitemid 32108530)
pfcrt Allelic types with two novel amino acid mutations in chloroquine resistant Plasmodium falciparum isolates from the Philippines. Chen N, Kyle DE, Pasay C, Fowler EV, Baker J, Peters JM, Cheng Q, Antimicrob Agents Chemother 2003 47 3500 3505 10.1128/AAC.47.11.3500-3505.2003 14576108 (Pubitemid 37345710)
A comparison of the in vitro activities of amodiaquine and desethylamodiaquine against isolates of Plasmodium falciparum. Childs GE, Boudreau EF, Milhous WK, Wimonwattratee T, Pooyindee N, Pang L, Davidson DE Jr, Am J Trop Med Hyg 1989 40 7 11 2644858 (Pubitemid 19063924)
Short report: association between chloroquine and amodiaquine resistance and allelic variation in the Plasmodium falciparum multiple drug resistance 1 gene and the chloroquine resistance transporter gene in isolates from the upper Nile in southern Sudan. Ochong EO, van den Broek IV, Keus K, Nzila A, Am J Trop Med Hyg 2003 69 184 187 13677373
Geographic patterns of Plasmodium falciparum drug resistance distinguished by differential responses to amodiaquine and chloroquine. Sa JM, Twu O, Hayton K, Reyes S, Fay MP, Ringwald P, Wellems TE, Proc Natl Acad Sci U S A 2009 106 18883 18889 10.1073/pnas.0911317106 19884511
Amodiaquine resistance in Plasmodium falciparum malaria in Afghanistan is associated with the pfcrt SVMNT allele at codons 72 to 76. Beshir K, Sutherland CJ, Merinopoulos I, Durrani N, Leslie T, Rowland M, Hallett RL, Antimicrob Agents Hemother 2010 54 3714 3716 10.1128/AAC.00358-10
Genetic diversity and chloroquine selective sweeps in Plasmodium falciparum. Wootton JC, Feng X, Ferdig MT, Cooper RA, Mu J, Baruch DI, Magill AJ, Su XZ, Nature 2002 418 320 323 10.1038/nature00813 12124623 (Pubitemid 34790681)
Evolution of a unique Plasmodium falciparum chloroquine-resistance phenotype in association with Pfcrt polymorphism in Papua New Guinea and South America. Mehlotra RK, Fujioka H, Roepe PD, Janneh O, Ursos LMB, Jacobs-Lorena V, McNamara DT, Bockarie MJ, Kazura JW, Kyle DE, Fidock DA, Zimmerman PA, Proc Natl Acad Sci U S A 2001 98 12689 12694 10.1073/pnas.221440898 11675500 (Pubitemid 33020007)
Occurrence of the Southeast Asian/South American SVMNT haplotype of the chloroquine-resistance transporter gene in Plasmodium falciparum in Tanzania. Alifrangis M, Dalgaard MB, Lusingu JP, Vestergaard LS, Staalsoe T, Jensen AT, Enevold A, Ronn AM, Khalil IF, Warhurst DC, Lemnge MM, Theander TG, Bygbjerg IC, J Infect Dis 2006 193 1738 1741 10.1086/504269 16703518 (Pubitemid 43839479)
Plasmodium falciparum isolates from Angola show the SVMNT haplotype in the pfcrt gene. Gama BE, de Carvalho GA P, LutucutaKosi FJ, de Oliveira NK A, Fortes F, Rosenthal PJ, Daniel Ribeiro CT, da Cruz MD F, Malar J 2010 9 174 10.1186/1475-2875-9-174 20565881
Assessing the efficacy of chloroquine and sulfadoxine-pyrimethamine for treatment of uncomplicated Plasmodium falciparum malaria in the Democratic Republic of Congo. Kazadi WM, Vong S, Makina BN, Mantshumba JC, Kabuya W, Kebela B, Ngimbi NP, Trop Med Int Health 2003 8 868 875 10.1046/j.1365-3156.2003. 01098.x 14516297 (Pubitemid 37305000)
Giemsa-stained thick blood films as a source of DNA for Plasmodium species-specific real-time PCR. Cnops L, Van Esbroeck M, Bottieau E, Jacobs J, Malar J 2010 9 370 10.1186/1475-2875-9-370 21176207
Validation of a four-primer real-time PCR as a diagnostic tool for single and mixed Plasmodium infections. Cnops L, Jacobs J, Van Esbroeck M, Clin Microbiol Infect 2011 17 1101 1107 10.1111/j.1469-0691.2010.03344.x 20718798
Molecular monitoring of Plasmodium falciparum drug susceptibility at the time of the introduction of artemisinin-based combination therapy in Yaoundé, Cameroon: implications for the future. Menard S, Morlais I, Tahar R, Sayang C, IssamouMayengue P, Iriart X, Benoit-Vical F, Lemen B, Magnaval J-F, Awono-Ambene P, Basco LK, Berry A, Malar J 2012 11 113 10.1186/1475-2875-11-113 22498364
Prevalence of pfcrt point mutations and level of chloroquine resistance in Plasmodium falciparum isolates from Africa. Severini C, Menegon M, Sannella AR, Paglia MG, Narciso P, Matteelli A, Gulletta M, Caramello P, Canta F, Xayavong MV, Moura IN, Pieniazek NJ, Taramelli D, Majori G, Infect Genet Evol 2006 6 262 268 10.1016/j.meegid.2005.07.002 16154388 (Pubitemid 43785812)
Prevalence of pfcrt mutations in Congolese and Malawian Plasmodium falciparum isolates as determined by a new Taqman assay. Wilson PE, Kazadi W, Kamwendo DD, Mwapasa V, Purfield A, Meshnick SR, Acta Trop 2005 93 97 106 10.1016/j.actatropica.2004.09.010 15589802 (Pubitemid 39611359)
Resistance-mediating Polymorphisms in Plasmodium falciparum infections in Kinshasa, Democratic Republic of the Congo. Mobula L, Lilley B, Tshefu AK, Rosenthal PJ, Am J Trop Med Hyg 2009 80 555 558 19346374
Reemergence of chloroquine-sensitive Plasmodium falciparum malaria after cessation of chloroquine in Malawi. Kublin JG, Cortese JF, Njunju EM, Mukadam RA, Wirima JJ, Kazembe PN, Djimdé AA, Kouriba B, Taylor TE, Plowe CV, J Infect Dis 2003 187 1870 1875 10.1086/375419 12792863 (Pubitemid 36724485)
Declining chloroquine resistance of Plasmodium falciparum in Lambarene, Gabon from 1992 to 1998. Schwenke A, Brandts C, Philipps J, Winkler S, Wernsdorfer WH, Kremsner PG, Wien Klin Wochenschr 2001 113 63 64 11233471 (Pubitemid 32120896)
Longitudinal study assessing the return of chloroquine susceptibility of Plasmodium falciparum in isolates from travellers returning from West and Central Africa, 2000-2011. Gharbi M, Flegg JA, Hubert V, Kendjo E, Metcalf JE, Bertaux L, Guérin PJ, Le Bras J, Members of the French National Reference Centre for Imported Malaria Study, Malar J 2013 12 35 10.1186/1475-2875-12-35 23351608
Population genetic analyses of Plasmodium falciparum chloroquine receptor transporter gene haplotypes reveal the evolutionary history of chloroquine-resistant malaria in India. Awasthi G, Prasad GBKS, Das A, Int J Parasitol 2011 41 705 709 10.1016/j.ijpara.2011.03.002 21447338
Are transporter genes other than the chloroquine resistance locus (pfcrt) and multidrug resistance gene (pfmdr) associated with antimalarial drug resistance? Anderson TJ, Nair S, Qin H, Singlam S, Brockman A, Paiphun L, Nosten F, Antimicrob Agents Chemother 2005 49 2180 2188 10.1128/AAC.49.6.2180-2188. 2005 15917511 (Pubitemid 40734444)
Selection of pfmdr1 mutations after amodiaquine monotherapy and amodiaquine plus artemisinin combination therapy in East Africa. Holmgren GJ, Hamrin J, Svard J, Mårtensson A, Gil JP, Bjorkman A, Infect Genet Evol 2007 7 562 569 10.1016/j.meegid.2007.03.005 17467344 (Pubitemid 47338282)
Resistance-mediating Plasmodium falciparum pfcrt and pfmdr1 alleles after treatment with artesunate-amodiaquine in Uganda. Nsobya SL, Dokomajilar C, Joloba M, Dorsey G, Rosenthal PJ, Antimicrob Agents Chemother 2007 51 3023 3025 10.1128/AAC.00012-07 17562798 (Pubitemid 47206253)
New haplotypes of the Plasmodium falciparum chloroquine resistance transporter (pfcrt) gene among chloroquine-resistant parasite isolates. Nagesha SH, Casey GJ, Rieckmann KH, Fryauff DJ, Laksana BS, Reeder JC, Maguire JD, Baird JK, Am J Trop Med Hyg 2003 68 398 402 12875286 (Pubitemid 38230637)
Patterns of chloroquine use and resistance in sub-Saharan Africa: a systematic review of household survey and molecular data. Frosch AEP, Venkatesan M, Laufer MK, Malar J 2011 10 11 10.1186/1475-2875-10-11 21235784
Protecting the malaria drug arsenal: halting the rise and spread of amodiaquine resistance by monitoring the PfCRT SVMNT type. Sa JM, Twu O, Malar J 2010 9 374 10.1186/1475-2875-9-374 21182787
In vitro amodiaquine resistance and its association with mutations in pfcrt and pfmdr1 genes of Plasmodium falciparum isolates from Nigeria. Folarin OA, Bustamante C, Gbotosho GO, Sowunmi A, Zalis MG, Oduola AMJ, Happi CT, Acta Trop 2011 120 224 230 10.1016/j.actatropica.2011.08.013 21920347
