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.
Department of Schistosomiasis and Endemic Diseases, Wuhan City Center for Disease Prevention and Control, Wuhan 430024, China.
Infect Genet Evol. 2022 Jul;101:105286. doi: 10.1016/j.meegid.2022.105286. Epub 2022 Apr 22.
Molecular markers for monitoring resistance could help improve malaria treatment policies. Delayed clearance of Plasmodium falciparum by artemisinin-based combination therapies (ACTs) has been reported in several countries. In addition to PfKelch13 (pfk13), new drug resistance genes, P. falciparum ubiquitin-specific protease 1 (pfubp1) and the eadaptor protein complex 2 mu subunit (pfap2mu), have been identified as being linked to ACTs. This study investigated the prevalence of single-nucleotide polymorphisms (SNPs) in clinical P. falciparum isolates pfubp1 and pfap2mu imported from Africa and Southeast Asia (SEA) to Wuhan, China, to provide baseline data for antimalarial resistance monitoring in this region.
Peripheral venous blood samples were collected in Wuhan, China, from August 2011 to December 2019. The Pfubp1 and pfap2mu SNPs of P. falciparum were determined by nested PCR and Sanger sequencing.
In total, 296 samples were collected. Subsequently, 92.23% (273/296) were successfully amplified and sequenced for Pfubp1. There were 60.07% (164/273) wild-type strains and 39.93% (109/273) mutant strains. The pfap2mu gene was divided into three fragments for amplification, and 82.77% (245/296), 90.20% (267/296) and 94.59% (280/296) were sequenced successfully. Genotypes reportedly associated with ACTs resistance detected in this study included pfubp1 D1525E as well as E1528D and pfap2mu S160N. The mutation prevalence rates were 10.99% (30/273), 13.19% (36/273) and 11.24% (30/267), respectively. These are all focused on Congo, Nigeria and Angola. Known delayed-clearance parasite mutations have also been found in SEA.
The existence of mutation sites of known clearance genes detected in the isolates in this study, including D1525E and E1528D in the pfubp1 gene and S160N in the pfap2mu gene, further proved the risk of ACTs resistance. Constant vigilance is therefore needed to protect the effectiveness of ACTs and to prevent the spread of drug-resistant P. falciparum. Further studies in malaria-endemic countries are needed to further validate potential genetic markers for monitoring parasite populations in Africa and SEA.
分子标志物可用于监测抗药性，有助于改善疟疾治疗政策。在几个国家都有报道称，青蒿素为基础的联合疗法（ACTs）延迟清除恶性疟原虫。除 PfKelch13（pfk13）外，新的耐药基因，疟原虫泛素特异性蛋白酶 1（pfubp1）和 e 衔接蛋白复合物 2 微结构域（pfap2mu），也被认为与 ACTs 有关。本研究调查了从非洲和东南亚（SEA）输入到中国武汉的临床疟原虫分离株 pfubp1 和 pfap2mu 中的单核苷酸多态性（SNPs）的流行情况，为该地区抗疟药耐药性监测提供了基线数据。
2011 年 8 月至 2019 年 12 月，在中国武汉采集外周静脉血样本。通过巢式 PCR 和 Sanger 测序确定恶性疟原虫 Pfubp1 和 pfap2mu 的 SNPs。
共采集 296 个样本。随后，92.23%（273/296）成功扩增并对 Pfubp1 进行测序。其中 60.07%（164/273）为野生型，39.93%（109/273）为突变型。pfap2mu 基因分为三个片段进行扩增，82.77%（245/296）、90.20%（267/296）和 94.59%（280/296）成功测序。本研究中检测到的与 ACTs 耐药相关的基因型包括 pfubp1 D1525E 以及 E1528D 和 pfap2mu S160N。突变率分别为 10.99%（30/273）、13.19%（36/273）和 11.24%（30/267）。这些都集中在刚果、尼日利亚和安哥拉。还在 SEA 发现了与延迟清除寄生虫相关的已知突变。
本研究中分离株检测到已知清除基因的突变位点，包括 pfubp1 基因中的 D1525E 和 E1528D 以及 pfap2mu 基因中的 S160N，进一步证明了 ACTs 耐药的风险。因此，需要持续保持警惕，以保护 ACTs 的有效性，并防止耐药性恶性疟原虫的传播。需要在疟疾流行国家进一步研究，以进一步验证非洲和 SEA 寄生虫群体监测的潜在遗传标记。
