SHORT COMMUNICATIONS

Analysis on the copy number variation of multidrug resistance-1 gene in 122 imported cases of falciparum malaria in Changsha

  • Bin TIAN ,
  • Yu LIAO ,
  • Lan WEN ,
  • Fang XIAO ,
  • Bin ZHANG ,
  • Xiao-jun SHEN
Expand
  • Changsha Center for Disease Control and Prevention, Changsha 410001, China

Received date: 2021-04-25

  Revised date: 2021-08-27

  Online published: 2022-01-12

Supported by

Natural Science Foundation of Hunan Province(2020JJ804)

Abstract

To understand the copy number variation (CNV)of Plasmodium falciparum multidrug resistence-1 (Pfmdr1) gene among imported Plasmodium falciparum, real-time fluorescence quantitative PCR was used to detect Pfmdr1 and β-tubulin (Pftub) gene from dried blood samples on filter paper collected from imported falciparum malaria cases in Changsha City from 2016 to 2019, with Pftub as the reference gene, for calculating the Pfmdr1 CNV in the samples examined. SPSS 23.0 statistic software was used to analyze the detection results and individuals’ clinical treatment information. The gene detection indicated that among 122 samples, 18 (14.8%) had Pfmdr1 CNV, with the variation rate 14.8% (18/122). The average of Pfmdr1 CNV in the samples collected from 2016 to 2019 was 1.020 ± 0.076, 1.136 ± 0.403, 1.387 ± 0.657 and 1.142 ± 0.349, respectively. The blood samples with CNV of the falciparum malaria cases were imported from Equatorial Guinea, Angola, Benin, Sierra Leone, Democratic Republic of Congo, Nigeria, Cameroon, Ghana and The Republic of Congo. The average Pfmdr1 CNV of the blood samples of the cases imported from East, West and Central Africa were 0.999 ± 0.073, 1.150 ± 0.368 and 1.249 ± 0.448, respectively, with significant difference between the regions (t = 2.663, 3.995, P < 0.05). The average medication duration for the cases with Pfmdr1 varied CNV was (5.93 ± 0.94) d, which was longer than that from non-varied CNV [(3.21 ± 1.23) d] (t = 8.930, P < 0.01). One month after medication, recrudescence occurred in two cases with varied CNV Pfmdr1, but in one case with non-varied CNV (likelyhood ratio χ 2 = 3.831, P < 0.05). One year after treatment ended, recrudescence occurred in two cases with varied CNV, whereas in one case with non-varied CNV (likelyhood ratio χ 2 = 5.372,P < 0.05). Gametocytes were detected in the blood smears of 5 cases with varies CNV, while in 3 cases with non-varied CNV (likelyhood ratio χ2 = 10.599, P < 0.01). Pfmdr1 gene CNV was found in the imported P. falciparum parasite in Changsha, and the variation may result in longer treatment duration for the patients and unable to completely clear up the parasites.

Cite this article

Bin TIAN , Yu LIAO , Lan WEN , Fang XIAO , Bin ZHANG , Xiao-jun SHEN . Analysis on the copy number variation of multidrug resistance-1 gene in 122 imported cases of falciparum malaria in Changsha[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2022 , 40(1) : 127 -131 . DOI: 10.12140/j.issn.1000-7423.2022.01.021

References

[1] Haldar K, Bhattacharjee S, Safeukui I. Drug resistance in Plasmodium[J]. Nat Rev Microbiol, 2018, 16(3): 156-170.
[2] Rout S, Mahapatra RK. Plasmodium falciparum: multidrug resistance[J]. Chem Biol Drug Des, 2019, 93(5): 737-759.
[3] Duah NO, Matrevi SA, de Souza DK, et al. Increased Pfmdr1 gene copy number and the decline in Pfcrt and Pfmdr1 resistance alleles in Ghanaian Plasmodium falciparum isolates after the change of anti-malarial drug treatment policy[J]. Malar J, 2013, 12: 377.
[4] Ye SY, Cheng YY, Li M, et al. Advances in methods for detecting drug-resistance molecular markers of Plasmodium falciparum[J]. Chin J Parasitol Parasit Dis, 2020, 38(4): 490-495. (in Chinese)
[4] (叶升玉, 成依依, 李曼, 等. 恶性疟原虫抗药性分子标记检测方法研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2020, 38(4): 490-495.)
[5] Basco LK, Le Bras J, Rhoades Z, et al. Analysis of Pfmdr1 and drug susceptibility in fresh isolates of Plasmodium falciparum from subsaharan Africa[J]. Mol Biochem Parasitol, 1995, 74(2): 157-166.
[6] Sidhu AB, Uhlemann AC, Valderramos SG, et al. Decreasing Pfmdr1 copy number in Plasmodium falciparum malaria heightens susceptibility to mefloquine, lumefantrine, halofantrine, quinine, and artemisinin[J]. J Infect Dis, 2006, 194(4): 528-535.
[7] Calçada C, Silva M, Baptista V, et al. Expansion of a specific Plasmodium falciparum Pfmdr1 haplotype in Southeast Asia with increased substrate transport[J]. mBio, 2020, 11(6): e02093-e02113.
[8] Labadie-Bracho M, Adhin MR. Increased Pfmdr1 copy number in Plasmodium falciparum isolates from Suriname[J]. Trop Med Int Health, 2013, 18(7): 796-799.
[9] Wilson CM, Volkman SK, Thaithong S, et al. Amplification of Pfmdr1 associated with mefloquine and halofantrine resistance in Plasmodium falciparum from Thailand[J]. Mol Biochem Parasitol, 1993, 57(1): 151-160.
[10] Venkatesan M, Gadalla NB, Stepniewska K, et al. Polymorphisms in Plasmodium falciparum chloroquine resistance transporter andmultidrug resistance 1 genes: parasite risk factors that affect treatment outcomes for P. falciparum malaria after artemether-lumefantrine and artesunate-amodiaquine[J]. Am J Trop Med Hyg, 2014, 91(4): 833-843.
[11] Tian B, Shen XJ, Liao Y, et al. Analysis of copy number variation in Pfpm2 gene of Plasmodium falciparum in 122 imported cases in Changsha from 2016 to 2019[J]. Acta Acad Med Milit Terti, 2020, 42(20): 2012-2017. (in Chinese)
[11] (田斌, 申晓君, 廖瑜, 等. 长沙市122例输入性恶性疟原虫Pfpm2基因拷贝数变异的分析[J]. 第三军医大学学报, 2020, 42(20): 2012-2017.)
[12] Price R, Robinson G, Brockman A, et al. Assessment of Pfmdr1 gene copy number by tandem competitive polymerase chain reaction[J]. Mol Biochem Parasitol, 1997, 85(2): 161-169.
[13] Win AA, Imwong M, Kyaw MP, et al. K13 mutations and Pfmdr1 copy number variation in Plasmodium falciparum malaria in Myanmar[J]. Malar J, 2016, 15: 297.
[14] Muhamad P, Chaijaroenkul W, Congpuong K, et al. SYBR greenⅠand TaqMan quantitative real-time polymerase chain reaction methods for the determination of amplification of Plasmodium falciparum multidrug resistance-1 gene (Pfmdr1)[J]. J Parasitol, 2011, 97(5): 939-942.
[15] Mungthin M, Watanatanasup E, Sitthichot N, et al. Influence of the Pfmdr1 gene on in vitro sensitivities of piperaquine in thai isolates of Plasmodium falciparum[J]. Am J Trop Med Hyg, 2017, 96(3): 624-629.
[16] Qidwai T. Exploration of copy number variation in genes related to anti-malarial drug resistance in Plasmodium falciparum[J]. Gene, 2020, 736: 144414.
[17] Ye SY, Cheng YY, Li M, et al. An overview on the resistance of Plasmodium falciparum to primaryanti-malarial drugs in China[J]. Chin J Parasitol Parasit Dis, 2020, 38(5): 631-636. (in Chinese)
[17] (叶升玉, 成依依, 李曼, 等. 我国恶性疟原虫主要药物抗性研究[J]. 中国寄生虫学与寄生虫病杂志, 2020, 38(5): 631-636.)
[18] Cowman AF, Galatis D, Thompson JK. Selection for mefloquine resistance in Plasmodium falciparum is linked to amplification of the Pfmdr1 gene and cross-resistance to halofantrine and quinine[J]. Proc Natl Acad Sci USA, 1994, 91(3): 1143-1147.
[19] Pacheco C, Moreno J, Herrera F. A high number of Pfmdr1 gene copies in P. falciparum from Venezuela[J]. Parasitol Res, 2019, 118(10): 3085-3089.
[20] Kiaco K, Teixeira J, Machado M, et al. Evaluation of artemether-lumefantrine efficacy in the treatment of uncomplicated malaria and its association with Pfmdr1, Pfatpase6 and K13-propeller polymorphisms in Luanda, Angola[J]. Malar J, 2015, 14: 504.
[21] Nguetse CN, Adegnika AA, Agbenyega T, et al. Molecular markers of anti-malarial drug resistance in central, west and east African children with severe malaria[J]. Malar J, 2017, 16: 217.
[22] Nyunt MH, Wang B, Aye KM, et al. Molecular surveillance of artemisinin resistance falciparum malaria among migrant goldmine workers in Myanmar[J]. Malar J, 2017, 16: 97.
[23] Ehrlich HY, Jones J, Parikh S. Molecular surveillance of antimalarial partner drug resistance in sub-Saharan Africa: a spatial-temporal evidence mapping study[J]. Lancet Microbe, 2020, 1(5): e209-e217.
[24] Yang B, Sun YF, Lei Y, et al. Research progress on the treatment of malaria with artemisinin andits derivatives[J]. Chin J Parasitol Parasit Dis, 2021, 39(3): 393-402. (in Chinese)
[24] (杨博, 孙毅凡, 雷瑶, 等. 青蒿素及其衍生物治疗疟疾的研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2021, 39(3): 393-402.)
Outlines

/

〈 〉