Objective To identify Plasmodium falciparum chloroquine resistant transporter (Pfcrt)point mutations and P. falciparum multidrug resistance 1 (pfmdr1)and their correlation in imported P. falciparum. Methods Blood samples were collected from returnees infected with P. falciparum in endemic areas of Africa and Southeast Asia during 2009-2016 in Wuhan city. The Pfcrt76 gene and pfmdr1 gene were amplified by nested PCR, and the products were digested by restriction endonuclease ApoⅠ, AseⅠ and EcoRv. The mutation rate was analyzed. Results A total of 232 patients with falciparum malaria were involved in the study. The mutation rates for Pfcrt76, pfmdr1 loci 86, 1042, and 1246 were 55.2%, 17.2%, 5.2%, and 8.6% respectively, the total mutation rate for pfmdr1 was 26.3%. The mutation rates for pfmdr1 loci 86, 1042, 1246 and pfmdr1 were 28.6%, 3.8%, 12.4%, and 36.2%, respectively in the samples with mutation in Pfcrt76, but 10.4%, 2.1%, 7.3%, and 17.7%, respectively in the samples without mutation in Pfcrt76 from Africa. The linkage disequilibrium analysis between symptoms and the mutations of Pfcrt76, pfmdr186, 1042, and 1246 were conducted, and the value of D'and r2 were 0.230 and 0.018, 0.290 and 0.004, 0.996 and 0.012, 0.150 and 0.035, respectively, for each mutation. Conclusion The pfmdr1 mutation sites between Africa and Southeast Asia have statistically significant differences. Pfcrt76 was positively correlated with pfmdr1 mutation and pfmdr186 point mutation.
ZHOU Shui-mao, LI Lian-jun, JIA Xi-shuai, YANG Yan, XU Ming-xing, CHEN Fang
. Correlation analysis on mutations of chloroquine resistant transporter and multi-drug resistance 1 gene in Plasmodium falciparum imported to Wuhan, China[J]. Chinese Journal of Vector Biology and Control, 2018
, 29(3)
: 242
-245
.
DOI: 10.11853/j.issn.1003.8280.2018.03.006
[1] Chandra RS,Orazem J,Ubben D,et al. Creative solutions to extraordinary challenges in clinical trials:methodology of a phase Ⅲ trial of azithromycin and chloroquine fixed-dose combination in pregnant women in Africa[J]. Malar J, 2013, 12:122.
[2] World Health Organization. World malaria report 2015[R]. Geneva:WHO, 2015.
[3] Mita T,Tanabe K. Evolution of Plasmodium falciparum drug resistance:implications for the development and containment of artemisinin resistance[J]. Jpn J Infect Dis,2012,65(6):465-475.
[4] Picot S,Olliaro P,de Monbrison F,et al. A systematic review and Meta-analysis of evidence for correlation between molecular markers of parasite resistance and treatment outcome in falciparum malaria[J]. Malar J, 2009, 8:89.
[5] Mungthin M,Suwandittakul N,Chaijaroenkul W,et al. The patterns of mutation and amplification of Plasmodium falciparum Pfcrt and pfmdr1 genes in Thailand during the year 1988 to 2003[J]. Parasitol Res, 2010, 107(3):539-545.
[6] Dajem SMB,Al-Qahtani A. Analysis of gene mutations involved in chloroquine resistance in Plasmodium falciparum parasites isolated from patients in the southwest of Saudi Arabia[J]. Ann Saudi Med, 2010, 30(3):187-192.
[7] Wellems TE, Panton LJ, Gluzman HY, et al. Chloroquine resistance not linked to mdr-like genes in a Plasmodium falciparum cross[J]. Nature, 1990, 345(6272):253-255.
[8] Wellems TE,Walker-Jonah A,Panton LJ. Genetic mapping of the chloroquine-resistance locus on Plasmodium falciparum chromosome 7[J]. Proc Natl Sci USA, 1991, 88(8):3382-3386.
[9] 汤林华. 输入性疟疾的诊治与管理[M]. 上海:上海科学技术出版社, 2010:30-45.
[10] 周水茂,杨燕,吴凯,等. 非洲输入性恶性疟病例裂殖子表面蛋白1等位基因型检测[J]. 中国寄生虫学与寄生虫病杂志, 2015, 33(4):247-250.
[11] 周水茂,杨燕,陈智,等. 输入性恶性疟原虫Pfcrt基因76位点多态性分析[J]. 中国血吸虫病防治杂志, 2013, 25(4):402-404.
[12] 张丽,丰俊,张少森,等. 2015年全国疟疾疫情分析[J]. 中国寄生虫学与寄生虫病杂志, 2016, 34(6):477-481.
[13] 郭传坤,黎军,林康明,等. 广西2010-2012年输入性疟疾流行病学分析[J]. 中国媒介生物学及控制杂志, 2014, 25(4):333-336.
[14] 王亚丽,王煊,张彦平. 2005-2013年全国本地感染与境外输入疟疾流行病学特征分析[J]. 中国媒介生物学及控制杂志, 2015, 26(2):120-126.
[15] 官亚宜,汤林华. 恶性疟原虫药物抗性相关的分子标记及其在药物抗性监测中的应用[J]. 中国寄生虫学与寄生虫病杂志, 2005, 23(2):117-120.
[16] 张逸龙,潘卫庆. 恶性疟原虫对青蒿素产生抗性的研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2015, 33(6):418-424.
[17] 王珊珊,董莹,邓艳,等. 中缅边境恶性疟原虫pfmdr1基因多态性及pfmsp1等位基因分型研究[J]. 国际医学寄生虫病杂志, 2012, 39(2):85-88.
[18] Lekana-Douki JB, Dinzouna Boutamba SD, Zatra R, et al. Increased prevalence of the Plasmodium falciparum pfmdr1 86N genotype among field isolates from Franceville,Gabon after replacement of chloroquine by artemether-lumefantrine and artesunate-mefloquine[J]. Infect Genet Evol, 2011, 11(2):512-517.
[19] Mawili-Mboumba DP,Kun JF,Lell B,et al. Pfmdr1 alleles and response to ultralow-dose mefloquine treatment in Gabonese patients[J]. Antimicrob Agents Chemother,2002,46(1):166-170.
[20] World Health Organization. Guidelines for the treatment of malaria[R]. Geneva:WHO, 2006.
[21] Mawili-Mboumba DP,Ngomo JMN,Maboko F,et al. Pfcrt 76T and pfmdr1 86Y allele frequency in Plasmodium falciparum isolates and use of self-medication in a rural area of Gabon[J]. Trans R Soc Trop Med Hyg, 2014, 108(11):729-734.
[22] Holmgren G,Hamrin J,Svärd J,et al. Selection of pfmdr1 mutations after amodiaquine monotherapy and amodiaquine plus artemisinin combination therapy in East Africa[J]. Infect Genet Evol, 2007, 7(5):562-569.
[23] Lobo E,de Sousa B,Rosa S,et al. Prevalence of pfmdr1 alleles associated with artemether-lumefantrine tolerance/resistance in Maputo before and after the implementation of artemisinin-based combination therapy[J]. Malar J, 2014, 13:300.
[24] Thomas SM,Ndir O,Dieng T,et al. In vitro chloroquine susceptibility and PCR analysis of Pfcrt and pfmdr1 polymorphisms in Plasmodium falciparum isolates from Senegal[J]. Am J Trop Med Hyg, 2002, 66(5):474-480.
[25] 徐超,魏庆宽,李瑾,等. 输入性恶性疟原虫耐药相关基因Pfcrt和pfmdr1单倍型及突变分析[J]. 中国人兽共患病学报, 2016, 32(12):1051-1057.
[26] 周瑞敏,王轶南,钱丹,等. 2015年河南省输入性恶性疟原虫Pfcrt基因的多态性分析[J]. 中国寄生虫学与寄生虫病杂志, 2016, 34(5):399-404.
[27] 邓艳,王珊珊,董莹,等. 中缅边境恶性疟原虫Pfcrt基因76位点突变研究[J]. 中国人兽共患病学报, 2014, 30(11):1137-1140.
