Skip to main content
Top
Published in: Malaria Journal 1/2020

01-12-2020 | Plasmodium Falciparum | Research

Molecular assessment of kelch13 non-synonymous mutations in Plasmodium falciparum isolates from Central African Republic (2017–2019)

Authors: Romaric Nzoumbou-Boko, Chris-Boris Gildas Panté-Wockama, Carine Ngoagoni, Nathalie Petiot, Eric Legrand, Ulrich Vickos, Jean-Chrysostome Gody, Alexandre Manirakiza, Christophe Ndoua, Jean-Pierre Lombart, Didier Ménard

Published in: Malaria Journal | Issue 1/2020

Login to get access

Abstract

Background

Over the last decade, artemisinin-based combination therapy (ACT) has contributed substantially to the decrease in malaria-related morbidity and mortality. The emergence of Plasmodium falciparum parasites resistant to artemisinin derivatives in Southeast Asia and the risk of their spread or of local emergence in sub-Saharan Africa are a major threat to public health. This study thus set out to estimate the proportion of P. falciparum isolates, with Pfkelch13 gene mutations associated with artemisinin resistance previously detected in Southeast Asia.

Methods

Blood samples were collected in two sites of Bangui, the capital of the Central African Republic (CAR) from 2017 to 2019. DNA was extracted and nested PCR were carried out to detect Plasmodium species and mutations in the propeller domain of the Pfkelch13 gene for P. falciparum samples.

Results

A total of 255 P. falciparum samples were analysed. Plasmodium ovale DNA was found in four samples (1.57%, 4/255). Among the 187 samples with interpretable Pfkelch13 sequences, four samples presented a mutation (2.1%, 4/187), including one non-synonymous mutation (Y653N) (0.5%, 1/187). This mutation has never been described as associated with artemisinin resistance in Southeast Asia and its in vitro phenotype is unknown.

Conclusion

This preliminary study indicates the absence of Pfkelch13 mutant associated with artemisinin resistance in Bangui. However, this limited study needs to be extended by collecting samples across the whole country along with the evaluation of in vitro and in vivo phenotype profiles of Pfkelch13 mutant parasites to estimate the risk of artemisinin resistance in the CAR.
Literature
1.
go back to reference Cibulskis ER, Alonso P, Aponte J, Aregawi M, Barrette A, Bergeron L. Malaria: global progress 2000–2015 and future challenges. Infect Dis Poverty. 2016;5:61.CrossRef Cibulskis ER, Alonso P, Aponte J, Aregawi M, Barrette A, Bergeron L. Malaria: global progress 2000–2015 and future challenges. Infect Dis Poverty. 2016;5:61.CrossRef
2.
go back to reference WHO. World malaria report. Geneva: World Health Organization; 2019. WHO. World malaria report. Geneva: World Health Organization; 2019.
3.
go back to reference Noedl H, Se Y, Schaecher K, Smith BL, Socheat D, Fukuda MM, et al. Evidence of artemisinin-resistant malaria in western Cambodia. N Engl J Med. 2008;359:2619–20.CrossRef Noedl H, Se Y, Schaecher K, Smith BL, Socheat D, Fukuda MM, et al. Evidence of artemisinin-resistant malaria in western Cambodia. N Engl J Med. 2008;359:2619–20.CrossRef
4.
go back to reference Hassett MR, Roepe PD. Origin and spread of evolving artemisinin-resistant Plasmodium falciparum malarial parasites in Southeast Asia. Am J Trop Med Hyg. 2019;101:1204–11.CrossRef Hassett MR, Roepe PD. Origin and spread of evolving artemisinin-resistant Plasmodium falciparum malarial parasites in Southeast Asia. Am J Trop Med Hyg. 2019;101:1204–11.CrossRef
5.
go back to reference Balikagala B, Mita T, Ikeda M, Sakurai M, Yatsushiro S, Takahashi N, et al. Absence of in vivo selection for K13 mutations after artemether–lumefantrine treatment in Uganda. Malar J. 2017;16:23.CrossRef Balikagala B, Mita T, Ikeda M, Sakurai M, Yatsushiro S, Takahashi N, et al. Absence of in vivo selection for K13 mutations after artemether–lumefantrine treatment in Uganda. Malar J. 2017;16:23.CrossRef
6.
go back to reference Djaman JA, Olefongo D, Ako AB, Roman J, Ngane VF, Basco LK, Tahar R. Molecular epidemiology of malaria in Cameroon and Côte d’Ivoire. XXXI. Kelch 13 propeller sequences in Plasmodium falciparum isolates before and after implementation of artemisinin-based combination therapy. Am J Trop Med Hyg. 2017;97:222–4.CrossRef Djaman JA, Olefongo D, Ako AB, Roman J, Ngane VF, Basco LK, Tahar R. Molecular epidemiology of malaria in Cameroon and Côte d’Ivoire. XXXI. Kelch 13 propeller sequences in Plasmodium falciparum isolates before and after implementation of artemisinin-based combination therapy. Am J Trop Med Hyg. 2017;97:222–4.CrossRef
7.
go back to reference Manirakiza A, Njuimo SP, Le Faou A, Malvy D, Millet P. Availability of antimalarial drugs and evaluation of the attitude and practices for the treatment of uncomplicated malaria in Bangui, Central African Republic. J Trop Med. 2010;2010:510834.CrossRef Manirakiza A, Njuimo SP, Le Faou A, Malvy D, Millet P. Availability of antimalarial drugs and evaluation of the attitude and practices for the treatment of uncomplicated malaria in Bangui, Central African Republic. J Trop Med. 2010;2010:510834.CrossRef
8.
go back to reference Menard D, Djalle D, Manirakiza A, Yapou F, Siadoua V, Sana S, et al. Drug-resistant malaria in Bangui, Central African Republic: an in vitro assessment. Am J Trop Med Hyg. 2005;73:239–43.CrossRef Menard D, Djalle D, Manirakiza A, Yapou F, Siadoua V, Sana S, et al. Drug-resistant malaria in Bangui, Central African Republic: an in vitro assessment. Am J Trop Med Hyg. 2005;73:239–43.CrossRef
9.
go back to reference Javelle E, Madamet M, Gaillard T, Velut G, Surcouf C, Michel R, et al. Delayed onset of Plasmodium falciparum malaria after doxycycline prophylaxis in a soldier returning from the Central African Republic. Antimicrob Agents Chemother. 2016;60:2592–3.CrossRef Javelle E, Madamet M, Gaillard T, Velut G, Surcouf C, Michel R, et al. Delayed onset of Plasmodium falciparum malaria after doxycycline prophylaxis in a soldier returning from the Central African Republic. Antimicrob Agents Chemother. 2016;60:2592–3.CrossRef
10.
go back to reference Nambei WS, Lango Yaya E, Pounguinza S, Achonduh O, Bogon A, Lengande R, et al. Efficacy and safety of antimalarial combinations for treatment of uncomplicated malaria in children in Bangui, Central African Republic. Med Sante Trop. 2013;23:313–9 (in French).PubMed Nambei WS, Lango Yaya E, Pounguinza S, Achonduh O, Bogon A, Lengande R, et al. Efficacy and safety of antimalarial combinations for treatment of uncomplicated malaria in children in Bangui, Central African Republic. Med Sante Trop. 2013;23:313–9 (in French).PubMed
11.
go back to reference Ménard D, Khim N, Beghain J, Adegnika AA, Shafiul-Alam M, Amodu O. A worldwide map of Plasmodium falciparum K13-propeller polymorphisms. N Engl J Med. 2016;374:2453–64.CrossRef Ménard D, Khim N, Beghain J, Adegnika AA, Shafiul-Alam M, Amodu O. A worldwide map of Plasmodium falciparum K13-propeller polymorphisms. N Engl J Med. 2016;374:2453–64.CrossRef
12.
go back to reference Ariey F, Witkowski B, Amaratunga C, Beghain J, Langlois AC, Khim N, et al. A molecular marker of artemisinin-resistant Plasmodium falciparum malaria. Nature. 2014;505:50–5.CrossRef Ariey F, Witkowski B, Amaratunga C, Beghain J, Langlois AC, Khim N, et al. A molecular marker of artemisinin-resistant Plasmodium falciparum malaria. Nature. 2014;505:50–5.CrossRef
13.
go back to reference WHO. Global technical strategy for malaria 2016–2030. Geneva: World Health Organization; 2016. WHO. Global technical strategy for malaria 2016–2030. Geneva: World Health Organization; 2016.
14.
go back to reference Walsh PS, Metzger DA, Higuchi R. Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques. 1991;10:506–13.PubMed Walsh PS, Metzger DA, Higuchi R. Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques. 1991;10:506–13.PubMed
15.
go back to reference Singh B, Bobogare A, Cox-Singh J, Snounou G, Abdullah MS, Rahman HA. A genus- and species-specific nested polymerase chain reaction malaria detection assay for epidemiologic studies. Am J Trop Med Hyg. 1999;60:687–92.CrossRef Singh B, Bobogare A, Cox-Singh J, Snounou G, Abdullah MS, Rahman HA. A genus- and species-specific nested polymerase chain reaction malaria detection assay for epidemiologic studies. Am J Trop Med Hyg. 1999;60:687–92.CrossRef
16.
go back to reference Djallé D, Gody JC, Moyen JM, Tekpa G, Ipero J, Madji N, et al. Performance of Paracheck™-Pf, SD Bioline malaria Ag-Pf and SD Bioline malaria Ag-Pf/pan for diagnosis of falciparum malaria in the Central African Republic. BMC Infect Dis. 2014;14:109.CrossRef Djallé D, Gody JC, Moyen JM, Tekpa G, Ipero J, Madji N, et al. Performance of Paracheck™-Pf, SD Bioline malaria Ag-Pf and SD Bioline malaria Ag-Pf/pan for diagnosis of falciparum malaria in the Central African Republic. BMC Infect Dis. 2014;14:109.CrossRef
17.
go back to reference Bichara C, Flahaut P, Costa D, Bienvenu AL, Picot S, Gargala G. Cryptic Plasmodium ovale concurrent with mixed Plasmodium falciparum and Plasmodium malariae infection in two children from Central African Republic. Malar J. 2017;16:339.CrossRef Bichara C, Flahaut P, Costa D, Bienvenu AL, Picot S, Gargala G. Cryptic Plasmodium ovale concurrent with mixed Plasmodium falciparum and Plasmodium malariae infection in two children from Central African Republic. Malar J. 2017;16:339.CrossRef
18.
go back to reference Guerra RI, Ore M, Valdivia HO, Bishop DK, Ramos M, Mores CN, Campbell WR. A cluster of the first reported Plasmodium ovale spp. infections in Peru occuring among returning UN peace-keepers, a review of epidemiology, prevention and diagnostic challenges in non-endemic regions. Malar J. 2019;18:176.CrossRef Guerra RI, Ore M, Valdivia HO, Bishop DK, Ramos M, Mores CN, Campbell WR. A cluster of the first reported Plasmodium ovale spp. infections in Peru occuring among returning UN peace-keepers, a review of epidemiology, prevention and diagnostic challenges in non-endemic regions. Malar J. 2019;18:176.CrossRef
19.
go back to reference WHO. Artemisinin resistance and artemisinin-based combination therapy efficacy: status report. Geneva: World Health Organization; 2018. WHO. Artemisinin resistance and artemisinin-based combination therapy efficacy: status report. Geneva: World Health Organization; 2018.
20.
go back to reference Yang C, Zhang H, Zhou R, Qian D, Liu Y, Zhao Y, et al. Polymorphisms of Plasmodium falciparum k13-propeller gene among migrant workers returning to Henan Province, China from Africa. BMC Infect Dis. 2017;17:560.CrossRef Yang C, Zhang H, Zhou R, Qian D, Liu Y, Zhao Y, et al. Polymorphisms of Plasmodium falciparum k13-propeller gene among migrant workers returning to Henan Province, China from Africa. BMC Infect Dis. 2017;17:560.CrossRef
21.
go back to reference Kamau E, Campino S, Amenga-Etego L, Drury E, Ishengoma D, Johnson K, et al. K13-propeller polymorphisms in Plasmodium falciparum parasites from sub-Saharan Africa. J Infect Dis. 2015;211:1352–5.PubMed Kamau E, Campino S, Amenga-Etego L, Drury E, Ishengoma D, Johnson K, et al. K13-propeller polymorphisms in Plasmodium falciparum parasites from sub-Saharan Africa. J Infect Dis. 2015;211:1352–5.PubMed
22.
go back to reference Ouattara A, Kone A, Adams M, Fofana B, Maiga AW, Hampton S. Polymorphisms in the K13-propeller gene in artemisinin-susceptible Plasmodium falciparum parasites from Bougoula-Hameau and Bandiagara, Mali. Am J Trop Med Hyg. 2015;92:1202–6.CrossRef Ouattara A, Kone A, Adams M, Fofana B, Maiga AW, Hampton S. Polymorphisms in the K13-propeller gene in artemisinin-susceptible Plasmodium falciparum parasites from Bougoula-Hameau and Bandiagara, Mali. Am J Trop Med Hyg. 2015;92:1202–6.CrossRef
23.
go back to reference Taylor SM, Parobek CM, DeConti DK, Kayentao K, Coulibaly SO, Greenwood BM, et al. Absence of putative artemisinin resistance mutations among Plasmodium falciparum in sub-Saharan Africa: a molecular epidemiologic study. J Infect Dis. 2015;211:680–8.CrossRef Taylor SM, Parobek CM, DeConti DK, Kayentao K, Coulibaly SO, Greenwood BM, et al. Absence of putative artemisinin resistance mutations among Plasmodium falciparum in sub-Saharan Africa: a molecular epidemiologic study. J Infect Dis. 2015;211:680–8.CrossRef
24.
go back to reference Lu F, Culleton R, Zhang M, Ramaprasad A, von Seidlein L, Zhou H, et al. Emergence of indigenous artemisinin-resistant Plasmodium falciparum in Africa. N Engl J Med. 2017;376:991–3.CrossRef Lu F, Culleton R, Zhang M, Ramaprasad A, von Seidlein L, Zhou H, et al. Emergence of indigenous artemisinin-resistant Plasmodium falciparum in Africa. N Engl J Med. 2017;376:991–3.CrossRef
25.
go back to reference Mishra N, Prajapati SK, Kaitholia K, Bharti RS, Srivastava B, Phookan S, et al. Surveillance of artemisinin resistance in Plasmodium falciparum in India using the kelch13 molecular marker. Antimicrob Agents Chemother. 2015;59:2548–53.CrossRef Mishra N, Prajapati SK, Kaitholia K, Bharti RS, Srivastava B, Phookan S, et al. Surveillance of artemisinin resistance in Plasmodium falciparum in India using the kelch13 molecular marker. Antimicrob Agents Chemother. 2015;59:2548–53.CrossRef
26.
go back to reference Mayengue PI, Niama RF, Kouhounina Batsimba D, Malonga-Massanga A, Louzolo I, Loukabou Bongolo NC, et al. No polymorphisms in K13-propeller gene associated with artemisinin resistance in Plasmodium falciparum isolated from Brazzaville, Republic of Congo. BMC Infect Dis. 2018;18:538.CrossRef Mayengue PI, Niama RF, Kouhounina Batsimba D, Malonga-Massanga A, Louzolo I, Loukabou Bongolo NC, et al. No polymorphisms in K13-propeller gene associated with artemisinin resistance in Plasmodium falciparum isolated from Brazzaville, Republic of Congo. BMC Infect Dis. 2018;18:538.CrossRef
27.
go back to reference Eboumbou Moukoko CE, Huang F, Nsango SE, Kojom Foko LP, Ebong SB, Epee Eboumbou P, et al. K-13 propeller gene polymorphisms isolated between 2014 and 2017 from Cameroonian Plasmodium falciparum malaria patients. PLoS ONE. 2019;14:e0221895.CrossRef Eboumbou Moukoko CE, Huang F, Nsango SE, Kojom Foko LP, Ebong SB, Epee Eboumbou P, et al. K-13 propeller gene polymorphisms isolated between 2014 and 2017 from Cameroonian Plasmodium falciparum malaria patients. PLoS ONE. 2019;14:e0221895.CrossRef
28.
go back to reference Day T, Huijben S, Read AF. Is selection relevant in the evolutionary emergence of drug resistance? Trends Microbiol. 2015;23:126–33.CrossRef Day T, Huijben S, Read AF. Is selection relevant in the evolutionary emergence of drug resistance? Trends Microbiol. 2015;23:126–33.CrossRef
29.
go back to reference Hastings IM. Complex dynamics and stability of resistance to antimalarial drugs. Parasitology. 2006;132:615–24.CrossRef Hastings IM. Complex dynamics and stability of resistance to antimalarial drugs. Parasitology. 2006;132:615–24.CrossRef
30.
go back to reference Oboh MA, Ndiaye D, Antony HA, Badiane AS, Singh US, Ali NA, et al. Status of artemisinin resistance in malaria parasite Plasmodium falciparum from molecular analyses of the kelch13 gene in Southwestern Nigeria. Biomed Res Int. 2018;2018:2305062.CrossRef Oboh MA, Ndiaye D, Antony HA, Badiane AS, Singh US, Ali NA, et al. Status of artemisinin resistance in malaria parasite Plasmodium falciparum from molecular analyses of the kelch13 gene in Southwestern Nigeria. Biomed Res Int. 2018;2018:2305062.CrossRef
31.
go back to reference Maïga-Ascofaré O, May J. Is the A578S single-nucleotide polymorphism in K13-propeller a marker of emerging resistance to artemisinin among Plasmodium falciparum in Africa? J Infect Dis. 2016;213:165–6.CrossRef Maïga-Ascofaré O, May J. Is the A578S single-nucleotide polymorphism in K13-propeller a marker of emerging resistance to artemisinin among Plasmodium falciparum in Africa? J Infect Dis. 2016;213:165–6.CrossRef
32.
go back to reference Ogouyèmi-Hounto A, Damien G, Deme AB, Ndam NT, Assohou C, Tchonlin D, et al. Lack of artemisinin resistance in Plasmodium falciparum in northwest Benin after 10 years of use of artemisinin-based combination therapy. Parasite. 2016;23:28.CrossRef Ogouyèmi-Hounto A, Damien G, Deme AB, Ndam NT, Assohou C, Tchonlin D, et al. Lack of artemisinin resistance in Plasmodium falciparum in northwest Benin after 10 years of use of artemisinin-based combination therapy. Parasite. 2016;23:28.CrossRef
Metadata
Title
Molecular assessment of kelch13 non-synonymous mutations in Plasmodium falciparum isolates from Central African Republic (2017–2019)
Authors
Romaric Nzoumbou-Boko
Chris-Boris Gildas Panté-Wockama
Carine Ngoagoni
Nathalie Petiot
Eric Legrand
Ulrich Vickos
Jean-Chrysostome Gody
Alexandre Manirakiza
Christophe Ndoua
Jean-Pierre Lombart
Didier Ménard
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/s12936-020-03264-y

Other articles of this Issue 1/2020

Malaria Journal 1/2020 Go to the issue