恶性疟原虫青蒿素耐药分子监测研究进展

doi:10.12140/j.issn.1000-7423.2025.02.017

中国寄生虫学与寄生虫病杂志 ›› 2025, Vol. 43 ›› Issue (2): 260-268.doi: 10.12140/j.issn.1000-7423.2025.02.017

恶性疟原虫青蒿素耐药分子监测研究进展

李飞¹()(), 刘耀宝¹^,²^,^*()(), 曹俊¹^,²

1 南京医科大学公共卫生学院，江苏南京 211166
2 国家卫生健康委员会寄生虫病预防和控制技术重点实验室，江苏省寄生虫与媒介控制技术重点实验室，江苏省医学重点实验室，江苏省血吸虫病防治研究所，江苏无锡 214064

收稿日期:2024-09-18 修回日期:2024-12-04 出版日期:2025-04-30 发布日期:2025-04-14
通讯作者: * 刘耀宝（ORCID：0000-0001-9616-8948），男，博士，副研究员，从事寄生虫病防控与研究。E-mail：yaobao0721@163.com
作者简介:李飞（ORCID：0009-0000-9682-399X），男，硕士研究生，从事寄生虫病分子流行病学研究。E-mail：lifei197192@163.com
基金资助:
国家自然科学基金(82320108014);国家自然科学基金(82372275);国家自然科学基金(81971967);江苏省科教能力提升工程(ZDXYS202207)

Researches progress on molecular surveillance of artemisinin resistance in Plasmodium falciparum

LI Fei¹()(), LIU Yaobao¹^,²^,^*()(), CAO Jun¹^,²

1 School of Public Health, Nanjing Medical University, Nanjing 211166, Jiangsu, China
2 National Health Commission Key Laboratory of Parasitic Disease Control and Prevention, Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology, Jiangsu Provincial Medical Key Laboratory of Jiangsu Institute of Parasitic Diseases, Wuxi 214064, Jiangsu, China

Received:2024-09-18 Revised:2024-12-04 Online:2025-04-30 Published:2025-04-14
Contact: * E-mail：yaobao0721@163.com
Supported by:
National Natural Science Foundation of China(82320108014);National Natural Science Foundation of China(82372275);National Natural Science Foundation of China(81971967);Jiangsu Province Capability Improvement Project through Science, Technology and Education(ZDXYS202207)

摘要/Abstract

摘要：

随着以青蒿素为基础的联合疗法在恶性疟治疗中的广泛应用，恶性疟原虫对青蒿素及其衍生物逐渐产生了部分耐药性，成为当前全球疟疾防控最严峻的挑战之一。近年来，恶性疟原虫青蒿素耐药分子机制研究和分子标记鉴定取得较大进展。目前已发现的恶性疟原虫青蒿素耐药相关基因有Kelch13（pfk13）、泛素特异性蛋白酶1（pfubp1）、衔接蛋白复合体2-μ亚基（pfap2μ）和冠状蛋白（pfcoronin）等。分子监测结果显示，恶性疟原虫青蒿素耐药相关突变在东南亚地区已普遍存在，非洲、南美洲和大洋洲也已陆续出现了独立起源的耐药突变虫株。高通量测序技术和分子检测技术的发展为恶性疟原虫青蒿素耐药分子监测提供了强大助力。本文就恶性疟原虫青蒿素耐药分子监测的研究进展作一综述，以期为恶性疟原虫青蒿素耐药监测和控制提供参考。

关键词: 恶性疟原虫, 青蒿素耐药性, 分子监测, pfk13

Abstract:

The widespread deployment of artemisinin-based combination therapies (ACTs) for falciparum malaria has led to emergence of partial resistance to artemisinin and its derivatives, which has become one of the greatest challenge in the global malaria control programme. Recently, great strides have been achieved in unraveling the molecular mechanism underlying the resistance of P. falciparum to artemisinin and identification of molecular markers. The currently identified P. falciparum artemisinin resistance-associated genes include kelch protein 13 (pfk13), ubiquitin carboxyl-terminal hydrolase 1 (pfubp1), AP-2 complex subunit mu (pfap2μ), and coronin (pfcoronin). Molecular surveillance data reveal widespread distribution of P. falciparum artemisinin resistance-associated gene mutations across Southeast Asia, and emergence of artemisinin-resistant P. falciparum strains of independent origin in Africa, South America and Oceania. Advances in high-throughput sequencing and molecular testing provide powerful tools for molecular monitoring of artemisinin resistance in P. falciparum. This review summarizes the advances in molecular surveillance of artemisinin resistance in P. falciparum, aiming to provide insights into surveillance and management of artemisinin-resistant P. falciparum.

Key words: Plasmodium falciparum, Artemisinin resistance, Molecular surveillance, pfk13

中图分类号:

R531.3

李飞, 刘耀宝, 曹俊. 恶性疟原虫青蒿素耐药分子监测研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2025, 43(2): 260-268.

LI Fei, LIU Yaobao, CAO Jun. Researches progress on molecular surveillance of artemisinin resistance in Plasmodium falciparum[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2025, 43(2): 260-268.

图/表 5

图1

表1

表2

表3

非洲国家pfk13主要耐药突变的突变频率

年份	调查地	样本量/份	pfk13突变频率/%	SNP突变频率/%				参考文献
年份	调查地	样本量/份	pfk13突变频率/%	C469Y	R561H	A675V	R622I	参考文献
2010	卢旺达胡耶	75	0	0	0	0	0	[57]
2014	卢旺达胡耶	81	2.5	0	0	0	0	[57]
2015	卢旺达胡耶	66	4.5	0	0	1.5	0	[57]
2019	卢旺达胡耶	66	12.1	0	4.5	1.5	0	[58]
2023	卢旺达胡耶	212	19.3	0	9.0	5.7	0	[59]
2013	卢旺达马萨卡和鲁胡哈	32	12.5	0	0	0	0	[60]
2014	卢旺达马萨卡和鲁胡哈	102	17.6	0	6.9	0	0	[60]
2015	卢旺达马萨卡和鲁胡哈	400	11.5	0.25	3.0	0	0	[60]
2014—2015	卢旺达马萨卡	257	-	0	7.4	0	0	[60]
2018	卢旺达马萨卡	51	21.6	0	19.6	0	0	[61]
2018	卢旺达鲁卡拉	82	28.1	0	21.9	0	0	[61]
2021	卢旺达鲁卡拉	135	31.9	0	23.5	6.4	0	[61]
2018	卢旺达	219	-	-	11.9	-	-	[5]
2018—2019	卢旺达基加利	73	30.1	0	21.9	0	0	[62]
1999—2004	乌干达	683	2.3	0	0	0	0	[63]
2012—2016	乌干达	716	3.6	0.14	0	0.14	0	[63]
2015	乌干达北部（古卢）	505	3.9	0	0	0	0	[65]
2017	乌干达北部（古卢）	87	9.2	0	0	9.2	0	[65]
2018	乌干达北部（古卢）	60	18.3	1.7	0	13.3	0	[65]
2019	乌干达北部（古卢）	93	20.4	4.3	0	11.8	0	[65]
2016	乌干达	326	4.6	1.2	0	0.6	0	[64]
2017	乌干达	419	4.8	2.1	0	2.4	0	[64]
2018	乌干达	500	5.6	2.6	0	1.4	0	[64]
2019	乌干达	762	7.6	2.7	0	4.3	0	[64]
2020	乌干达	1 181	17.8	9.4	0	6.5	0	[64]
2021	乌干达	933	17.3	4.8	1.2	5.9	0	[64]
2022	乌干达	940	27.6	9.6	1.4	10.1	0	[64]
2016	厄立特里亚	280	9.7	0	0.3	0	8.6	[66]
2017	厄立特里亚	211	8.6	0	0	0	7.6	[66]
2018	厄立特里亚	-	-	-	-	-	16.7	[5]
2019	厄立特里亚	361	22.8	0	0	0	21.1	[66]
2014	埃塞俄比亚阿姆哈拉	125	2.4	0	0	0	2.4	[67]
2018	埃塞俄比亚阿姆哈拉	598	-	-	-	-	9.8	[39]
2012—2015	赤道几内亚	224	6.7	0	0	0	0	[69]
2013	赤道几内亚	1	-	-	-	-	-	[69]
2016	坦桑尼亚	320	5.3	0	0	0	0	[70]
2017	坦桑尼亚	774	-	0	0.2	0	0	[71]
2019	坦桑尼亚	422	7.3	0	0.2	0	0	[72]
2020	坦桑尼亚	96	4.2	0	0	0	1.1	[73]
2022	坦桑尼亚	173	-	-	22.5	-	-	[74]
2010—2015	刚果（金）	66	0	0	0	0	0	[75]
2017	刚果（金）	717	1.0	0	0	0	0	[76]
2018—2019	刚果（金）	325	2.2	0	0	0	0	[77]
2020—2021	刚果（金）	1 065	3.2	0	0.094	0	0	[78]

表3

表4

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方法	优点	缺点
PCR + RFLP	技术成熟，操作简便	灵敏度和特异性相对较低，难以检测低丰度突变
qPCR	特异性强，灵敏度高，结果快速准确	需要昂贵的荧光定量设备
ddPCR	高灵敏度，高特异性，可以对低丰度目标进行定量	成本较高，设备和技术要求严格
等温扩增技术	简便快速，不需要复杂设备，适合现场检测	难以对多个靶标进行扩增，限制了其在多个耐药基因检测中的应用
微阵列芯片	可以同时检测多个耐药基因SNP，高通量	定制化芯片成本高，技术要求高
一代测序（Sanger测序）	读长较长，准确度高，金标准	测序通量低，成本较高
二代测序	高通量，可同时测序大量DNA片段，效率高	数据处理复杂，成本较高
三代测序	长读长，适用于复杂基因组分析	成本高，技术相对较新，需要进一步优化

恶性疟原虫青蒿素耐药分子监测研究进展

Researches progress on molecular surveillance of artemisinin resistance in Plasmodium falciparum

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年份	调查地	样本量/份	pfk13突变频率/%	SNP突变频率/%					参考文献
年份	调查地	样本量/份	pfk13突变频率/%	C580Y	I543T	P553L	R539T	P574L	参考文献
2001—2002	柬埔寨拜林	40	> 70.0	45.0	-	-	-	-	[9]
2011—2012	柬埔寨拜林	84	> 95.0	95.0	-	-	-	-	[9]
2000—2005	越南	80	28.6	3.5	11.8	8.2	0	0	[44]
2006—2010	越南	84	39.3	4.8	31.0	0	1.2	0	[44]
2011—2012	越南	248	61.2	3.2	37.9	3.2	0.8	0	[44]
2013—2014	越南	126	46.8	14.3	6.3	7.9	8.7	0	[44]
2015—2016	越南	73	79.5	56.2	1.4	6.8	0	0	[44]
2017—2018	越南	171	65.5	58.5	0	4.7	0	0	[45]
2018—2019	越南	127	70.1	68.5	0	0	1.6	0	[46]
2002—2004	泰国西部	89	0	0	0	0	0	0	[49]
2004	泰国拉农府	48	20.8	12.5	0	2.1	0	6	[49]
2004	泰国攀牙湾	44	6.8	2.3	0	0	0	0	[49]
2012	泰国攀牙湾	4	25.0	0	0	0	0	0	[51]
2015	泰国攀牙湾	10	20.0	20.0	0	0	0	0	[51]
2005	泰国达叻	57	52.6	42.1	0	0	7.0	1.8	[49]
2003	泰国西部（泰缅边境）	43	12.5	0	0	0	0	0	[54]
2014	泰国西部（泰缅边境）	80	90.0	65.0	0	2.5	0	0	[54]
2012	泰国拉农府	8	25.0	12.0	0	0	0	0	[51]
2015	泰国拉农府	19	31.6	31.6	0	0	0	0	[51]
2016	泰国拉农府	9	100	66.7	0	0	33.3	0	[52]
2017	泰国也拉	12	0	0	0	0	0	0	[51]
2013	泰国（泰柬边境）	27	100	63.3	0	0	33.3	0	[50]
2014	泰国（泰柬边境）	36	100	82.1	0	0	17.9	0	[50]
2015	泰国（泰柬边境）	46	100	100	0	0	0	0	[50]