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Published in: Malaria Journal 1/2020

Open Access 01-12-2020 | Chloroquin | Research

Molecular surveillance of anti-malarial drug resistance in Democratic Republic of Congo: high variability of chloroquinoresistance and lack of amodiaquinoresistance

Authors: Doudou M. Yobi, Nadine K. Kayiba, Dieudonné M. Mvumbi, Raphael Boreux, Pius Z. Kabututu, Hippolyte N. T. Situakibanza, Joris L. Likwela, Patrick De Mol, Emile W. Okitolonda, Niko Speybroeck, Georges L. Mvumbi, Marie-Pierre Hayette

Published in: Malaria Journal | Issue 1/2020

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Abstract

Background

The loss of chloroquine (CQ) effectiveness has led to its withdrawal from national policies as a first-line treatment for uncomplicated malaria in several endemic countries, such as the Democratic Republic of Congo (DRC). The K76T mutation on the pfcrt gene has been identified as a marker of CQ resistance and the SVMNT haplotype in codons 72–76 on the same gene has been associated with resistance to amodiaquine (AQ). In the DRC, the prevalence of K76T has decreased from 100% in 2000 to 63.9% in 2014. The purpose of this study was to determine the prevalence of K76T mutations in circulating strains of Plasmodium falciparum, 16 years after CQ withdrawal in the DRC and to investigate the presence of the SVMNT haplotype.

Methods

In 2017, ten geographical sites across the DRC were selected. Dried blood samples were collected from patients attending health centres. Malaria was first detected by a rapid diagnostic test (RDT) available on site (SD Bioline Malaria Ag Pf or CareStart Malaria Pf) or thick blood smear and then confirmed by a P. falciparum species-specific real-time PCR assay. A pfcrt gene segment containing a fragment that encodes amino acids at positions 72–76 was amplified by conventional PCR before sequencing.

Results

A total of 1070 patients were enrolled. Of the 806 PCR-confirmed P. falciparum positive samples, 764 were successfully sequenced. The K76T mutation was detected in 218 samples (28.5%; 95% CI 25.4%–31.9%), mainly (96%) with the CVIET haplotype. Prevalence of CQ resistance marker was unequally distributed across the country, ranging from 1.5% in Fungurume to 89.5% in Katana. The SVMNT haplotype, related to AQ resistance, was not detected.

Conclusion

Overall, the frequency of the P. falciparum CQ resistance marker has decreased significantly and no resistance marker to AQ was detected in the DRC in 2017. However, the between regions variability of CQ resistance remains high in the country. Further studies are needed for continuous monitoring of the CQ resistance level for its prospective re-use in malaria management. The absence of the AQ resistance marker is in line with the use of this drug in the current DRC malaria treatment policy.
Literature
1.
go back to reference OMS. Stratégie technique mondiale de lutte contre le Paludisme 2016–2030. Genève: Organisation Mondiale de la Santé; 2015. OMS. Stratégie technique mondiale de lutte contre le Paludisme 2016–2030. Genève: Organisation Mondiale de la Santé; 2015.
2.
go back to reference Kazadi WM, Vong S, Makina BN, Mantshumba JC, Kabuya W, Kebela BI, et al. Assessing the efficacy of chloroquine and sulfadoxine-pyrimethamine for treatment of uncomplicated Plasmodium falciparum malaria in the Democratic Republic of Congo. Trop Med Int Health. 2003;8:868–75.CrossRef Kazadi WM, Vong S, Makina BN, Mantshumba JC, Kabuya W, Kebela BI, et al. Assessing the efficacy of chloroquine and sulfadoxine-pyrimethamine for treatment of uncomplicated Plasmodium falciparum malaria in the Democratic Republic of Congo. Trop Med Int Health. 2003;8:868–75.CrossRef
4.
go back to reference Martin RE. The malaria parasite’s chloroquine resistance transporter is a member of the drug/metabolite transporter superfamily. Mol Biol Evol. 2004;21:1938–49.CrossRef Martin RE. The malaria parasite’s chloroquine resistance transporter is a member of the drug/metabolite transporter superfamily. Mol Biol Evol. 2004;21:1938–49.CrossRef
5.
go back to reference Tran CV. The principal chloroquine resistance protein of Plasmodium falciparum is a member of the drug/metabolite transporter superfamily. Microbiology. 2004;150:1–3.CrossRef Tran CV. The principal chloroquine resistance protein of Plasmodium falciparum is a member of the drug/metabolite transporter superfamily. Microbiology. 2004;150:1–3.CrossRef
6.
go back to reference Ecker A, Lehane AM, Clain J, Fidock DA. PfCRT and its role in antimalarial drug resistance. Trends Parasitol. 2012;28:504–14.CrossRef Ecker A, Lehane AM, Clain J, Fidock DA. PfCRT and its role in antimalarial drug resistance. Trends Parasitol. 2012;28:504–14.CrossRef
7.
go back to reference Lakshmanan V, Bray PG, Verdier-Pinard D, Johnson DJ, Horrocks P, Muhle RA, et al. A critical role for PfCRT K76T in Plasmodium falciparum verapamil-reversible chloroquine resistance. EMBO J. 2005;24:2294–305.CrossRef Lakshmanan V, Bray PG, Verdier-Pinard D, Johnson DJ, Horrocks P, Muhle RA, et al. A critical role for PfCRT K76T in Plasmodium falciparum verapamil-reversible chloroquine resistance. EMBO J. 2005;24:2294–305.CrossRef
8.
go back to reference Cui L, Mharakurwa S, Ndiaye D, Rathod PK, Rosenthal PJ. Antimalarial drug resistance: literature review and activities and findings of the ICEMR network. Am J Trop Med Hyg. 2015;93:57–68.CrossRef Cui L, Mharakurwa S, Ndiaye D, Rathod PK, Rosenthal PJ. Antimalarial drug resistance: literature review and activities and findings of the ICEMR network. Am J Trop Med Hyg. 2015;93:57–68.CrossRef
9.
go back to reference 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;344:257–63.CrossRef 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;344:257–63.CrossRef
10.
go back to reference Awasthi G, Satya Prasad GBK, Satya GBK, Das A. Pfcrt haplotypes and the evolutionary history of chloroquine-resistant Plasmodium falciparum. Mem Inst Oswaldo Cruz. 2012;107:129–34.CrossRef Awasthi G, Satya Prasad GBK, Satya GBK, Das A. Pfcrt haplotypes and the evolutionary history of chloroquine-resistant Plasmodium falciparum. Mem Inst Oswaldo Cruz. 2012;107:129–34.CrossRef
11.
go back to reference Beshir K, Sutherland CJ, Merinopoulos I, Durrani N, Leslie T, Rowland M, et al. Amodiaquine Resistance in Plasmodium falciparum malaria in Afghanistan is associated with the pfcrt SVMNT allele at codons 72 to 76. Antimicrob Agents Chemother. 2010;54:3714–6.CrossRef Beshir K, Sutherland CJ, Merinopoulos I, Durrani N, Leslie T, Rowland M, et al. Amodiaquine Resistance in Plasmodium falciparum malaria in Afghanistan is associated with the pfcrt SVMNT allele at codons 72 to 76. Antimicrob Agents Chemother. 2010;54:3714–6.CrossRef
12.
go back to reference Alifrangis M, Dalgaard MB, Lusingu JP, Vestergaard LS, Staalsoe T, Jensen ATR, et al. Occurrence of the Southeast Asian/South American SVMNT haplotype of the chloroquine-resistance transporter gene in Plasmodium falciparum in Tanzania. J Infect Dis. 2006;193:1738–41.CrossRef Alifrangis M, Dalgaard MB, Lusingu JP, Vestergaard LS, Staalsoe T, Jensen ATR, et al. Occurrence of the Southeast Asian/South American SVMNT haplotype of the chloroquine-resistance transporter gene in Plasmodium falciparum in Tanzania. J Infect Dis. 2006;193:1738–41.CrossRef
13.
go back to reference Gama BE, Pereira-Carvalho GA, Lutucuta Kosi FJ, de Oliveira NK, Fortes F, Rosenthal PJ, et al. Plasmodium falciparum isolates from Angola show the SVMNT haplotype in the pfcrt gene. Malar J. 2010;9:174.CrossRef Gama BE, Pereira-Carvalho GA, Lutucuta Kosi FJ, de Oliveira NK, Fortes F, Rosenthal PJ, et al. Plasmodium falciparum isolates from Angola show the SVMNT haplotype in the pfcrt gene. Malar J. 2010;9:174.CrossRef
14.
go back to reference Kateera F, Nsobya SL, Tukwasibwe S, Hakizimana E, Mutesa L, Mens PF, et al. Molecular surveillance of Plasmodium falciparum drug resistance markers reveals partial recovery of chloroquine susceptibility but sustained sulfadoxine-pyrimethamine resistance at two sites of different malaria transmission intensities in Rwanda. Acta Trop. 2016;164:329–36.CrossRef Kateera F, Nsobya SL, Tukwasibwe S, Hakizimana E, Mutesa L, Mens PF, et al. Molecular surveillance of Plasmodium falciparum drug resistance markers reveals partial recovery of chloroquine susceptibility but sustained sulfadoxine-pyrimethamine resistance at two sites of different malaria transmission intensities in Rwanda. Acta Trop. 2016;164:329–36.CrossRef
15.
go back to reference Koukouikila-Koussounda F, Jeyaraj S, Nguetse CN, Nkonganyi CN, Kokou KC, Etoka-Beka MK, et al. Molecular surveillance of Plasmodium falciparum drug resistance in the Republic of Congo: four and nine years after the introduction of artemisinin-based combination therapy. Malar J. 2017;16:155.CrossRef Koukouikila-Koussounda F, Jeyaraj S, Nguetse CN, Nkonganyi CN, Kokou KC, Etoka-Beka MK, et al. Molecular surveillance of Plasmodium falciparum drug resistance in the Republic of Congo: four and nine years after the introduction of artemisinin-based combination therapy. Malar J. 2017;16:155.CrossRef
16.
go back to reference Lu F, Zhang M, Culleton RL, Xu S, Tang J, Zhou H, et al. Return of chloroquine sensitivity to Africa? Surveillance of African Plasmodium falciparum chloroquine resistance through malaria imported to China. Parasit Vectors. 2017;10:355.CrossRef Lu F, Zhang M, Culleton RL, Xu S, Tang J, Zhou H, et al. Return of chloroquine sensitivity to Africa? Surveillance of African Plasmodium falciparum chloroquine resistance through malaria imported to China. Parasit Vectors. 2017;10:355.CrossRef
17.
go back to reference Kiarie WC, Wangai L, Agola E, Kimani FT, Hungu C. Chloroquine sensitivity: diminished prevalence of chloroquine-resistant gene marker pfcrt-76 13 years after cessation of chloroquine use in Msambweni, Kenya. Malar J. 2015;14:328.CrossRef Kiarie WC, Wangai L, Agola E, Kimani FT, Hungu C. Chloroquine sensitivity: diminished prevalence of chloroquine-resistant gene marker pfcrt-76 13 years after cessation of chloroquine use in Msambweni, Kenya. Malar J. 2015;14:328.CrossRef
18.
go back to reference Afoakwah R, Boampong JN, Egyir-Yawson A, Nwaefuna EK, Verner ON, Asare KK. High prevalence of PfCRT K76T mutation in Plasmodium falciparum isolates in Ghana. Acta Trop. 2014;136:32–6.CrossRef Afoakwah R, Boampong JN, Egyir-Yawson A, Nwaefuna EK, Verner ON, Asare KK. High prevalence of PfCRT K76T mutation in Plasmodium falciparum isolates in Ghana. Acta Trop. 2014;136:32–6.CrossRef
19.
go back to reference Golassa L, Enweji N, Erko B, Aseffa A, Swedberg G. High prevalence of pfcrt-CVIET haplotype in isolates from asymptomatic and symptomatic patients in south-central Oromia, Ethiopia. Malar J. 2014;13:120.CrossRef Golassa L, Enweji N, Erko B, Aseffa A, Swedberg G. High prevalence of pfcrt-CVIET haplotype in isolates from asymptomatic and symptomatic patients in south-central Oromia, Ethiopia. Malar J. 2014;13:120.CrossRef
20.
go back to reference Meshnick SR, Janko M, Tshefu AK, Taylor SM, Emch M, Antonia AL. A Cross-sectional survey of Plasmodium falciparum pfcrt mutant haplotypes in the Democratic Republic of Congo. Am J Trop Med Hyg. 2014;90:1094–7.CrossRef Meshnick SR, Janko M, Tshefu AK, Taylor SM, Emch M, Antonia AL. A Cross-sectional survey of Plasmodium falciparum pfcrt mutant haplotypes in the Democratic Republic of Congo. Am J Trop Med Hyg. 2014;90:1094–7.CrossRef
21.
go back to reference Mvumbi DM, Boreux R, Sacheli R, Lelo M, Lengu B, Nani-Tuma S, et al. Assessment of pfcrt 72–76 haplotypes eight years after chloroquine withdrawal in Kinshasa, Democratic Republic of Congo. Malar J. 2013;12:459.CrossRef Mvumbi DM, Boreux R, Sacheli R, Lelo M, Lengu B, Nani-Tuma S, et al. Assessment of pfcrt 72–76 haplotypes eight years after chloroquine withdrawal in Kinshasa, Democratic Republic of Congo. Malar J. 2013;12:459.CrossRef
22.
go back to reference Mvumbi DM, Bobanga TL, Kayembe J-MN, Mvumbi GL, Situakibanza HN-T, Benoit-Vical F, et al. Molecular surveillance of Plasmodium falciparum resistance to artemisinin-based combination therapies in the Democratic Republic of Congo. PLoS One. 2017;12:e0179142.CrossRef Mvumbi DM, Bobanga TL, Kayembe J-MN, Mvumbi GL, Situakibanza HN-T, Benoit-Vical F, et al. Molecular surveillance of Plasmodium falciparum resistance to artemisinin-based combination therapies in the Democratic Republic of Congo. PLoS One. 2017;12:e0179142.CrossRef
23.
go back to reference Manguin S, Carnevale P, Mouchet J, Coosemans M, Julvez J, Richard-Lenoble D, et al. Biodiversity of malaria in the World. Paris: John Libbey Eurotext; 2008. p. 36. Manguin S, Carnevale P, Mouchet J, Coosemans M, Julvez J, Richard-Lenoble D, et al. Biodiversity of malaria in the World. Paris: John Libbey Eurotext; 2008. p. 36.
24.
go back to reference Cnops L, Jacobs J, Esbroeck MV. Validation of a four-primer real-time PCR as a diagnostic tool for single and mixed Plasmodium infections. Clin Microbiol Infect. 2011;17:1101–7.CrossRef Cnops L, Jacobs J, Esbroeck MV. Validation of a four-primer real-time PCR as a diagnostic tool for single and mixed Plasmodium infections. Clin Microbiol Infect. 2011;17:1101–7.CrossRef
25.
go back to reference Laufer MK, Thesing PC, Eddington ND, Masonga R, Dzinjalamala FK, Takala SL, et al. Return of chloroquine antimalarial efficacy in Malawi. N Engl J Med. 2006;355:1959–66.CrossRef Laufer MK, Thesing PC, Eddington ND, Masonga R, Dzinjalamala FK, Takala SL, et al. Return of chloroquine antimalarial efficacy in Malawi. N Engl J Med. 2006;355:1959–66.CrossRef
26.
go back to reference Mohammed A, Ndaro A, Kalinga A, Manjurano A, Mosha JF, Mosha DF, et al. Trends in chloroquine resistance marker, Pfcrt-K76T mutation ten years after chloroquine withdrawal in Tanzania. Malar J. 2013;12:415.CrossRef Mohammed A, Ndaro A, Kalinga A, Manjurano A, Mosha JF, Mosha DF, et al. Trends in chloroquine resistance marker, Pfcrt-K76T mutation ten years after chloroquine withdrawal in Tanzania. Malar J. 2013;12:415.CrossRef
27.
go back to reference Hemming-Schroeder E, Umukoro E, Lo E, Fung B, Tomás-Domingo P, Zhou G, et al. Impacts of antimalarial drugs on Plasmodium falciparum drug resistance markers, Western Kenya, 2003–2015. Am J Trop Med Hyg. 2018;98:692–9.CrossRef Hemming-Schroeder E, Umukoro E, Lo E, Fung B, Tomás-Domingo P, Zhou G, et al. Impacts of antimalarial drugs on Plasmodium falciparum drug resistance markers, Western Kenya, 2003–2015. Am J Trop Med Hyg. 2018;98:692–9.CrossRef
28.
go back to reference Mwanza S, Joshi S, Nambozi M, Chileshe J, Malunga P, Kabuya J-BB, et al. The return of chloroquine-susceptible Plasmodium falciparum malaria in Zambia. Malar J. 2016;15:584.CrossRef Mwanza S, Joshi S, Nambozi M, Chileshe J, Malunga P, Kabuya J-BB, et al. The return of chloroquine-susceptible Plasmodium falciparum malaria in Zambia. Malar J. 2016;15:584.CrossRef
29.
go back to reference Frosch AE, Venkatesan M, Laufer MK. Patterns of chloroquine use and resistance in sub-Saharan Africa: a systematic review of household survey and molecular data. Malar J. 2011;10:116.CrossRef Frosch AE, Venkatesan M, Laufer MK. Patterns of chloroquine use and resistance in sub-Saharan Africa: a systematic review of household survey and molecular data. Malar J. 2011;10:116.CrossRef
30.
go back to reference Laufer M, Plowe C. Withdrawing antimalarial drugs: impact on parasite resistance and implications for malaria treatment policies. Drug Resist Updates. 2004;7:279–88.CrossRef Laufer M, Plowe C. Withdrawing antimalarial drugs: impact on parasite resistance and implications for malaria treatment policies. Drug Resist Updates. 2004;7:279–88.CrossRef
31.
go back to reference Mobula L, Lilley B, Tshefu AK, Rosenthal PJ. Resistance-mediating polymorphisms in Plasmodium falciparum infections in Kinshasa, Democratic Republic of the Congo. Am J Trop Med Hyg. 2009;80:555–8.CrossRef Mobula L, Lilley B, Tshefu AK, Rosenthal PJ. Resistance-mediating polymorphisms in Plasmodium falciparum infections in Kinshasa, Democratic Republic of the Congo. Am J Trop Med Hyg. 2009;80:555–8.CrossRef
32.
go back to reference Republic Democratic of Congo. Demographic and Health Survey II 2013–2014. p 204. Republic Democratic of Congo. Demographic and Health Survey II 2013–2014. p 204.
33.
go back to reference Bushman M, Antia R, Udhayakumar V, de Roode JC. Within-host competition can delay evolution of drug resistance in malaria. PLoS Biol. 2018;16:e2005712.CrossRef Bushman M, Antia R, Udhayakumar V, de Roode JC. Within-host competition can delay evolution of drug resistance in malaria. PLoS Biol. 2018;16:e2005712.CrossRef
34.
go back to reference Wongsrichanalai C, Pickard AL, Wernsdorfer WH, Meshnick SR. Epidemiology of drug-resistant malaria. Lancet Infect Dis. 2002;2:209–18.CrossRef Wongsrichanalai C, Pickard AL, Wernsdorfer WH, Meshnick SR. Epidemiology of drug-resistant malaria. Lancet Infect Dis. 2002;2:209–18.CrossRef
35.
go back to reference Onori E. The problem of Plasmodium falciparum drug resistance in Africa south of the Sahara. Bull World Health Organ. 1984;62:55–62.PubMedPubMedCentral Onori E. The problem of Plasmodium falciparum drug resistance in Africa south of the Sahara. Bull World Health Organ. 1984;62:55–62.PubMedPubMedCentral
36.
go back to reference Delacollette C, Embonga B, Malengreau M. Response to chloroquine of infections with Plasmodium falciparum in the Kivu region of Zaïre. Preliminary observations. Ann Soc Belge Med Trop. 1983;63:171–3. Delacollette C, Embonga B, Malengreau M. Response to chloroquine of infections with Plasmodium falciparum in the Kivu region of Zaïre. Preliminary observations. Ann Soc Belge Med Trop. 1983;63:171–3.
Metadata
Title
Molecular surveillance of anti-malarial drug resistance in Democratic Republic of Congo: high variability of chloroquinoresistance and lack of amodiaquinoresistance
Authors
Doudou M. Yobi
Nadine K. Kayiba
Dieudonné M. Mvumbi
Raphael Boreux
Pius Z. Kabututu
Hippolyte N. T. Situakibanza
Joris L. Likwela
Patrick De Mol
Emile W. Okitolonda
Niko Speybroeck
Georges L. Mvumbi
Marie-Pierre Hayette
Publication date
01-12-2020
Publisher
BioMed Central
Published in
Malaria Journal / Issue 1/2020
Electronic ISSN: 1475-2875
DOI
https://doi.org/10.1186/s12936-020-03192-x

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