对输入性非洲恶性疟原虫 pfcrt、pfmdr1 和 pfk13 多态性的抗疟耐药性进行分子监测。
Molecular surveillance of anti-malarial resistance pfcrt, pfmdr1, and pfk13 polymorphisms in African Plasmodium falciparum imported parasites to Wuhan, China.
机构信息
Department of Human Parasitology, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, 442000, China.
Department of Infectious Diseases, Renmin Hospital, Hubei University of Medicine, Shiyan, 442000, China.
出版信息
Malar J. 2021 May 1;20(1):209. doi: 10.1186/s12936-021-03737-8.
BACKGROUND
Imported malaria parasites with anti-malarial drug resistance (ADR) from Africa is a serious public health challenge in non-malarial regions, including Wuhan, China. It is crucial to assess the ADR status in African Plasmodium falciparum isolates from imported malaria cases, as this will provide valuable information for rational medication and malaria control.
METHODS
During 2017-2019, a cross-sectional study was carried out in Wuhan, China. Peripheral blood 3 ml of returned migrant workers from Africa was collected. The target fragments from pfcrt, pfmdr1, and k13 propeller (pfk13) genes were amplified, sequenced, and analysed.
RESULTS
In total, 106 samples were collected. Subsequently, 98.11% (104/106), 100% (106/106), and 86.79% (92/106) of these samples were successfully amplified and sequenced for the pfcrt (72-76), pfmdr1, and pfk13 genes, respectively. The prevalence of the pfcrt 76 T, pfmdr1 86Y, and pfmdr1 184F mutations was 9.62, 4.72, and 47.17%, respectively. At codons 72-76, the pfcrt locus displayed three haplotypes, CVMNK (wild-type), CVIET (mutation type), CV M/I N/E K/T (mixed type), with 87.50%, 9.62%, and 2.88% prevalence, respectively. For the pfmdr1 gene, NY (wild type), NF and YF (mutant type), N Y/F, Y Y/F, and N/Y Y/F (mixed type) accounted for 34.91, 43.40, 3.77, 15.09, 0.94, and 1.89% of the haplotypes, respectively. A total of 83 isolates with six unique haplotypes were found in pfcrt and pfmdr1 combined haplotypes, of which NY-CVMNK and NF-CVMNK accounted for 40.96% (34/83) and 43.37% (36/83), respectively. Furthermore, 90 cases were successfully sequenced (84.91%, 90/106) at loci 93, 97, 101, and 145, and 78 cases were successfully sequenced (73.58%, 78/106) at loci 343, 353, and 356 for pfcrt. However, the mutation was observed only in locus 356 with 6.41%. For pfk13, mutations reported in Southeast Asia (at loci 474, 476, 493, 508, 527, 533, 537, 539, 543, 553, 568, 574, 578, and 580) and Africa (at loci 550, 561, 575, 579, and 589) were not observed.
CONCLUSIONS
The present data from pfcrt and pfmdr1 demonstrate that anti-malarial drugs including chloroquine, amodiaquine, and mefloquine, remain effective against malaria treatment in Africa. The new mutations in pfcrt related to piperaquine resistance remain at relatively low levels. Another source of concern is the artemether-lumefantrine resistance-related profiles of N86 and 184F of pfmdr1. Although no mutation in pfk13 is detected, molecular surveillance must continue.
背景
来自非洲的具有抗疟药物耐药性（ADR）的输入性疟原虫对包括中国武汉在内的非疟区是一个严重的公共卫生挑战。评估来自输入性疟疾病例的非洲恶性疟原虫分离株的 ADR 状况至关重要，因为这将为合理用药和疟疾控制提供有价值的信息。
方法
2017-2019 年，在中国武汉进行了一项横断面研究。采集来自非洲的归国农民工外周血 3ml。扩增、测序和分析 pfcr t、pfmdr1 和 k13 螺旋桨（pfk13）基因的靶片段。
结果
共采集 106 份样本。随后，分别成功扩增和测序了 pfcrt（72-76）、pfmdr1 和 pfk13 基因的 98.11%（104/106）、100%（106/106）和 86.79%（92/106）的样本。pfcrt 76T、pfmdr1 86Y 和 pfmdr1 184F 突变的流行率分别为 9.62%、4.72%和 47.17%。在 72-76 位密码子处，pfcrt 基因座显示三种单倍型，CVMNK（野生型）、CVIET（突变型）、CV M/I N/E K/T（混合型），流行率分别为 87.50%、9.62%和 2.88%。对于 pfmdr1 基因，NY（野生型）、NF 和 YF（突变型）、NY/F、YY/F 和 NY/Y Y/F（混合型）分别占 34.91%、43.40%、3.77%、15.09%、0.94%和 1.89%的单倍型。在 pfcrt 和 pfmdr1 联合单倍型中发现了 83 个具有六个独特单倍型的分离株，其中 NY-CVMNK 和 NF-CVMNK 分别占 40.96%（34/83）和 43.37%（36/83）。此外，在 pfcrt 的 93、97、101 和 145 位以及 343、353 和 356 位成功测序了 90 例（84.91%，90/106），在 pfcrt 的 474、476、493、508、527、533、537、539、543、553、568、574、578 和 580 位以及非洲的 550、561、575、579 和 589 位成功测序了 78 例（73.58%，78/106）。然而，只在 568 位观察到突变，突变率为 6.41%。在 pfk13 中，未观察到与东南亚（474、476、493、508、527、533、537、539、543、553、568、574、578 和 580 位）和非洲（550、561、575、579 和 589 位）相关的突变。
结论
来自 pfcrt 和 pfmdr1 的数据表明，包括氯喹、阿莫地喹和甲氟喹在内的抗疟药物对非洲的疟疾治疗仍然有效。与哌喹耐药相关的 pfcrt 新突变仍处于较低水平。另一个令人担忧的问题是 pfmdr1 的青蒿素-咯萘啶耐药相关的 N86 和 184F 突变。尽管未检测到 pfk13 的突变，但必须继续进行分子监测。
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