1. National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Chinese Center for Tropical Diseases Research, WHO Collaborating Center for Tropical Diseases, National Centre for International Research on Tropical Diseases, NHC Key Laboratory of Parasite and Vector Biology (National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention), Shanghai 200025, China 2. Yunnan Institute of Parasitic Diseases, Puer 665000, China 3. Yingjiang County for Disease Control and Prevention, Yingjiang 679300, China 4. Hainan Center for Disease Control & Prevention, Haikou 570203, China
The dihydrofolate reductase (dhfr) and dihydropteroate synthetase (dhps) genes of Plasmodium vivax, as antifolate resistance-associated genes were used for drug resistance surveillance. A total of 375 P. vivax isolates collected from different geographical locations in China in 2009–2019 were used to sequence Pvdhfr and Pvdhps. The majority of the isolates harbored a mutant type allele for Pvdhfr (94.5%) and Pvdhps (68.2%). The most predominant point mutations were S117T/N (77.7%) in Pvdhfr and A383G (66.8%) in Pvdhps. Amino acid changes were identified at nine residues in Pvdhfr. A quadruple-mutant haplotype at 57, 58, 61, and 117 was the most frequent (57.4%) among 16 distinct Pvdhfr haplotypes. Mutations in Pvdhps were detected at six codons, and the double-mutant A383G/A553G was the most prevalent (39.3%). Pvdhfr exhibited a higher mutation prevalence and greater diversity than Pvdhps in China. Most isolates from Yunnan carried multiple mutant haplotypes, while the majority of samples from temperate regions and Hainan Island harbored the wild type or single mutant type. This study indicated that the antifolate resistance levels of P. vivax parasites were different across China and molecular markers could be used to rapidly monitor drug resistance. Results provided evidence for updating national drug policy and treatment guidelines.
RN Price, RJ Commons, KE Battle, K Thriemer, K Mendis. Plasmodium vivax in the era of the shrinking P. falciparum map. Trends Parasitol 2020; 36(6): 560–570 https://doi.org/10.1016/j.pt.2020.03.009
pmid: 32407682
2
World Health Organization. World malaria report 2020. Geneva: WHO, 2020
3
HW Zhang, Y Liu, SS Zhang, BL Xu, WD Li, JH Tang, SS Zhou, F Huang. Preparation of malaria resurgence in China: case study of vivax malaria re-emergence and outbreak in Huang-Huai Plain in 2006. Adv Parasitol 2014; 86: 205–230 https://doi.org/10.1016/B978-0-12-800869-0.00008-1
pmid: 25476886
4
S Zhang, S Guo, X Feng, A Afelt, R Frutos, S Zhou, S Manguin. Anopheles vectors in mainland China while approaching malaria elimination. Trends Parasitol 2017; 33(11): 889–900 https://doi.org/10.1016/j.pt.2017.06.010
pmid: 28734898
5
SS Zhou, F Huang, JJ Wang, SS Zhang, YP Su, LH Tang. Geographical, meteorological and vectorial factors related to malaria re-emergence in Huang-Huai River of central China. Malar J 2010; 9(1): 337 https://doi.org/10.1186/1475-2875-9-337
pmid: 21092326
6
A Nzila. The past, present and future of antifolates in the treatment of Plasmodium falciparum infection. J Antimicrob Chemother 2006; 57(6): 1043–1054 https://doi.org/10.1093/jac/dkl104
pmid: 16617066
7
World Health Organization. Implementing malaria in pregnancy programs in the context of World Health Organization recommendations on antenatal care for a positive pregnancy experience. Geneva: WHO, 2018
8
World Health Organization. Intermittent preventive treatment for infants using sulfadoxinepyrimethamine (SP-IPTi) for malaria control in Africa: Implementation field guide. Geneva: WHO, 2011
9
World Health Organization. World malaria report 2019. Geneva: WHO, 2019
10
NS Mohamed, H Abdelbagi, HA Osman, AE Ahmed, AM Yousif, YB Edris, EY Osman, AR Elsadig, EE Siddig, M Mustafa, AA Mohammed, Y Ali, MM Osman, MS Ali, RA Omer, A Ahmed, CH Sibley. A snapshot of Plasmodium falciparum malaria drug resistance markers in Sudan: a pilot study. BMC Res Notes 2020; 13(1): 512 https://doi.org/10.1186/s13104-020-05363-0
pmid: 33160417
11
B Huang, S Huang, XZ Su, X Tong, J Yan, H Li, F Lu. Molecular surveillance of pvdhfr, pvdhps, and pvmdr-1 mutations in Plasmodium vivax isolates from Yunnan and Anhui provinces of China. Malar J 2014; 13(1): 346 https://doi.org/10.1186/1475-2875-13-346
pmid: 25179752
12
F Huang, S Zhou, S Zhang, W Li, H Zhang. Monitoring resistance of Plasmdium vivax: point mutations in dihydrofolate reductase gene in isolates from Central China. Parasit Vectors 2011; 4(1): 80 https://doi.org/10.1186/1756-3305-4-80
pmid: 21586132
13
National Health Commission of the People’s Republic of China. Technical regulations for application of antimalarials (WS/T 485-2016). 2016
14
M Imwong, S Pukrittakayamee, S Looareesuwan, G Pasvol, J Poirreiz, NJ White, G Snounou. Association of genetic mutations in Plasmodium vivax dhfr with resistance to sulfadoxine-pyrimethamine: geographical and clinical correlates. Antimicrob Agents Chemother 2001; 45(11): 3122–3127 https://doi.org/10.1128/AAC.45.11.3122-3127.2001
pmid: 11600366
15
M Imwong, S Pukrittayakamee, L Rénia, F Letourneur, JP Charlieu, U Leartsakulpanich, S Looareesuwan, NJ White, G Snounou. Novel point mutations in the dihydrofolate reductase gene of Plasmodium vivax: evidence for sequential selection by drug pressure. Antimicrob Agents Chemother 2003; 47(5): 1514–1521 https://doi.org/10.1128/AAC.47.5.1514-1521.2003
pmid: 12709316
16
P Thongdee, J Kuesap, K Rungsihirunrat, P Tippawangkosol, M Mungthin, K Na-Bangchang. Distribution of dihydrofolate reductase (dhfr) and dihydropteroate synthase (dhps) mutant alleles in Plasmodium vivax isolates from Thailand. Acta Trop 2013; 128(1): 137–143 https://doi.org/10.1016/j.actatropica.2013.07.005
pmid: 23880285
17
K Tantiamornkul, T Pumpaibool, J Piriyapongsa, R Culleton, U Lek-Uthai. The prevalence of molecular markers of drug resistance in Plasmodium vivax from the border regions of Thailand in 2008 and 2014. Int J Parasitol Drugs Drug Resist 2018; 8(2): 229–237 https://doi.org/10.1016/j.ijpddr.2018.04.003
pmid: 29677637
18
PE de Pécoulas, R Tahar, P Yi, KH Thai, LK Basco. Genetic variation of the dihydrofolate reductase gene in Plasmodium vivax in Snoul, northeastern Cambodia. Acta Trop 2004; 92(1): 1–6 https://doi.org/10.1016/j.actatropica.2004.03.011
pmid: 15301969
19
PB Asih, SS Marantina, R Nababan, NF Lobo, IE Rozi, W Sumarto, RM Dewi, S Tuti, AS Taufik, Mulyanto, RW Sauerwein, D Syafruddin. Distribution of Plasmodium vivax pvdhfr and pvdhps alleles and their association with sulfadoxine-pyrimethamine treatment outcomes in Indonesia. Malar J 2015; 14(1): 365 https://doi.org/10.1186/s12936-015-0903-0
pmid: 26395428
20
S Joy, SK Ghosh, RN Achur, DC Gowda, N Surolia. Presence of novel triple mutations in the pvdhfr from Plasmodium vivax in Mangaluru city area in the southwestern coastal region of India. Malar J 2018; 17(1): 167 https://doi.org/10.1186/s12936-018-2316-3
pmid: 29661235
21
K Rakmark, GR Awab, J Duanguppama, U Boonyuen, AM Dondorp, M Imwong. Polymorphisms in Plasmodium vivax antifolate resistance markers in Afghanistan between 2007 and 2017. Malar J 2020; 19(1): 251 https://doi.org/10.1186/s12936-020-03319-0
pmid: 32664924
22
C Barnadas, M Tichit, C Bouchier, A Ratsimbasoa, L Randrianasolo, R Raherinjafy, M Jahevitra, S Picot, D Ménard. Plasmodium vivax dhfr and dhps mutations in isolates from Madagascar and therapeutic response to sulphadoxine-pyrimethamine. Malar J 2008; 7(1): 35 https://doi.org/10.1186/1475-2875-7-35
pmid: 18302746
23
C Barnadas, L Timinao, S Javati, J Iga, E Malau, C Koepfli, LJ Robinson, N Senn, B Kiniboro, L Rare, JC Reeder, PM Siba, PA Zimmerman, H Karunajeewa, TM Davis, I Mueller. Significant geographical differences in prevalence of mutations associated with Plasmodium falciparum and Plasmodium vivax drug resistance in two regions from Papua New Guinea. Malar J 2015; 14(1): 399 https://doi.org/10.1186/s12936-015-0879-9
pmid: 26452541
24
G Snounou, S Viriyakosol, XP Zhu, W Jarra, L Pinheiro, VE do Rosario, S Thaithong, KN Brown. High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction. Mol Biochem Parasitol 1993; 61(2): 315–320 https://doi.org/10.1016/0166-6851(93)90077-B
pmid: 8264734
25
AL Price, NJ Patterson, RM Plenge, ME Weinblatt, NA Shadick, D Reich. Principal components analysis corrects for stratification in genome-wide association studies. Nat Genet 2006; 38(8): 904–909 https://doi.org/10.1038/ng1847
pmid: 16862161
KD Miller, HO Lobel, M Pappaioanou, LC Patchen, FC Churchill. Failures of combined chloroquine and Fansidar prophylaxis in American travelers to East Africa. J Infect Dis 1986; 154(4): 689–691 https://doi.org/10.1093/infdis/154.4.689
pmid: 3528321
28
CV Plowe, JF Cortese, A Djimde, OC Nwanyanwu, WM Watkins, PA Winstanley, JG Estrada-Franco, RE Mollinedo, JC Avila, JL Cespedes, D Carter, OK Doumbo. Mutations in Plasmodium falciparum dihydrofolate reductase and dihydropteroate synthase and epidemiologic patterns of pyrimethamine-sulfadoxine use and resistance. J Infect Dis 1997; 176(6): 1590–1596 https://doi.org/10.1086/514159
pmid: 9395372
29
MD Hastings, KM Porter, JD Maguire, I Susanti, W Kania, MJ Bangs, CH Sibley, JK Baird. Dihydrofolate reductase mutations in Plasmodium vivax from Indonesia and therapeutic response to sulfadoxine plus pyrimethamine. J Infect Dis 2004; 189(4): 744–750 https://doi.org/10.1086/381397
pmid: 14767830
30
World Health Organization. Methods for the surveillance of antimalarial drug efficacy. Geneva: WHO, 2009
31
A Shaukat, Q Ali, T Connelley, MAU Khan, MA Saleem, M Evans, I Rashid, ND Sargison, U Chaudhry. Selective sweep and phylogenetic models for the emergence and spread of pyrimethamine resistance mutations in Plasmodium vivax. Infect Genet Evol 2019; 68:221–230 https://doi.org/10.1016/j.meegid.2018.12.032
pmid: 30594654
32
U Leartsakulpanich, M Imwong, S Pukrittayakamee, NJ White, G Snounou, W Sirawaraporn, Y Yuthavong. Molecular characterization of dihydrofolate reductase in relation to antifolate resistance in Plasmodium vivax. Mol Biochem Parasitol 2002; 119(1): 63–73 https://doi.org/10.1016/S0166-6851(01)00402-9
pmid: 11755187
33
BK Na, HW Lee, SU Moon, TS In, K Lin, M Maung, GT Chung, JK Lee, TS Kim, Y Kong. Genetic variations of the dihydrofolate reductase gene of Plasmodium vivax in Mandalay Division, Myanmar. Parasitol Res 2005; 96(5): 321–325 https://doi.org/10.1007/s00436-005-1364-0
pmid: 15924223
34
Z Zhou. Malaria Control and Research in China. Beijing: People’s Medical Publishing House, 1991
35
A Yaqoob, AA Khattak, MF Nadeem, H Fatima, G Mbambo, A Ouattara, M Adams, N Zeeshan, S Takala-Harrison. Prevalence of molecular markers of sulfadoxine-pyrimethamine and artemisinin resistance in Plasmodium falciparum from Pakistan. Malar J 2018; 17(1): 471 https://doi.org/10.1186/s12936-018-2620-y
pmid: 30558587
36
PE de Pécoulas, R Tahar, T Ouatas, A Mazabraud, LK Basco. Sequence variations in the Plasmodium vivax dihydrofolate reductase-thymidylate synthase gene and their relationship with pyrimethamine resistance. Mol Biochem Parasitol 1998; 92(2): 265–273 https://doi.org/10.1016/S0166-6851(97)00247-8
pmid: 9657331
37
F Lu, CS Lim, DH Nam, K Kim, K Lin, TS Kim, HW Lee, JH Chen, Y Wang, J Sattabongkot, ET Han. Mutations in the antifolate-resistance-associated genes dihydrofolate reductase and dihydropteroate synthase in Plasmodium vivax isolates from malaria-endemic countries. Am J Trop Med Hyg 2010; 83(3): 474–479 https://doi.org/10.4269/ajtmh.2010.10-0004
pmid: 20810806
38
M Korsinczky, K Fischer, N Chen, J Baker, K Rieckmann, Q Cheng. Sulfadoxine resistance in Plasmodium vivax is associated with a specific amino acid in dihydropteroate synthase at the putative sulfadoxine-binding site. Antimicrob Agents Chemother 2004; 48(6): 2214–2222 https://doi.org/10.1128/AAC.48.6.2214-2222.2004
pmid: 15155224
39
M Miao, Z Yang, L Cui, J Ahlum, Y Huang, L Cui. Different allele prevalence in the dihydrofolate reductase and dihydropteroate synthase genes in Plasmodium vivax populations from China. Am J Trop Med Hyg 2010; 83(6): 1206–1211 https://doi.org/10.4269/ajtmh.2010.10-0259
pmid: 21118923
40
LY Shang, CG Xue, SZ Su. Evaluation of the effect of 40 years anti-malaria measure in Henan Province. Chin J Parasitol Parasit Dis (Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi) 2000; 18(3): 189 (in Chinese)
pmid: 12567710
41
XZ Liu, BL Xu. Malaria situation and evaluation on the control effect in Henan Province during 1990−2005. Chin J Parasitol Parasit Dis (Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi) 2006; 24(3): 226–229(in Chinese) PMID:17094630
42
HL Yang. Retrospect and prospect of application of antimalarial drugs in Yunnan Province. J Practical Parasitic Dis 1999; 7(4): 174–176
43
VN Hawkins, H Joshi, K Rungsihirunrat, K Na-Bangchang, CH Sibley. Antifolates can have a role in the treatment of Plasmodium vivax. Trends Parasitol 2007; 23(5): 213–222 https://doi.org/10.1016/j.pt.2007.03.002
pmid: 17368986
44
D Kyabayinze, A Cattamanchi, MR Kamya, PJ Rosenthal, G Dorsey. Validation of a simplified method for using molecular markers to predict sulfadoxine-pyrimethamine treatment failure in African children with falciparum malaria. Am J Trop Med Hyg 2003; 69(3): 247–252 https://doi.org/10.4269/ajtmh.2003.69.247
pmid: 14628939
45
T Triglia, AF Cowman. Primary structure and expression of the dihydropteroate synthetase gene of Plasmodium falciparum. Proc Natl Acad Sci USA 1994; 91(15): 7149–7153 https://doi.org/10.1073/pnas.91.15.7149
pmid: 8041761
46
AM Brashear, AC Huckaby, Q Fan, LJ Dillard, Y Hu, Y Li, Y Zhao, Z Wang, Y Cao, J Miao, JL Guler, L Cui. New Plasmodium vivax genomes from the China−Myanmar border. Front Microbiol 2020; 11: 1930 https://doi.org/10.3389/fmicb.2020.01930
pmid: 32849480
47
Y Liu, S Auburn, J Cao, H Trimarsanto, H Zhou, KA Gray, TG Clark, RN Price, Q Cheng, R Huang, Q Gao. Genetic diversity and population structure of Plasmodium vivax in Central China. Malar J 2014; 13(1): 262 https://doi.org/10.1186/1475-2875-13-262
pmid: 25008859
48
YC Li, GZ Wang, F Meng, W Zeng, CH He, XM Hu, SQ Wang. Genetic diversity of Plasmodium vivax population before elimination of malaria in Hainan Province, China. Malar J 2015; 14(1): 78 https://doi.org/10.1186/s12936-015-0545-2
pmid: 25888891
49
B Huang, S Huang, XZ Su, H Guo, Y Xu, F Xu, X Hu, Y Yang, S Wang, F Lu. Genetic diversity of Plasmodium vivax population in Anhui Province of China. Malar J 2014; 13(1): 13 https://doi.org/10.1186/1475-2875-13-13
pmid: 24401153
50
F Huang, S Zhou, S Zhang, H Zhang, W Li. Meteorological factors-based spatio-temporal mapping and predicting malaria in central China. Am J Trop Med Hyg 2011; 85(3): 560–567 https://doi.org/10.4269/ajtmh.2011.11-0156
pmid: 21896823
51
S Ding, R Ye, D Zhang, X Sun, H Zhou, TF McCutchan, W Pan. Anti-folate combination therapies and their effect on the development of drug resistance in Plasmodium vivax. Sci Rep 2013; 3(1): 1008 https://doi.org/10.1038/srep01008
pmid: 23301149
52
JY Pan, SS Zhou, X Zheng, F Huang, DQ Wang, YZ Shen, YP Su, GC Zhou, F Liu, JJ Jiang. Vector capacity of Anopheles sinensis in malaria outbreak areas of central China. Parasit Vectors 2012; 5(1): 136 https://doi.org/10.1186/1756-3305-5-136
pmid: 22776520
53
World Health Organization. Artemisinin resistance and artemisinin-based combination therapy efficacy. Geneva: WHO, 2019