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Abstract：
BACKGROUND: Monitoring therapeutic efficacy is important to ensure the efficacy of artemisinin-based combination therapy (ACT) for malaria. The current first-line treatment for uncomplicated malaria recommended by the National Malaria Control Program in Niger is artemether-lumefantrine (AL). In 2020, an in vivo study was carried out to evaluate clinical and parasitological responses to AL as well as the molecular resistance to the drug in three sentinel sites: Agadez, Tessaoua and Gaya, in Niger.
METHODS: A multi-center, single-arm trial was conducted according to the 28-day World Health Organization (WHO) 2009 therapeutic efficacy study protocol. Children between 6 months and 15 years with confirmed uncomplicated Plasmodium falciparum infection and 1000-200,000 asexual parasites/μL of blood were enrolled and followed up for 28 days. Uncorrected and PCR-corrected efficacy results at day 28 were calculated, and molecular correction was performed by genotyping the msp1, msp2, and glurp genes. The pfk13, pfdhfr, pfdhps, pfcrt and pfmdr genes were analyzed by PCR and Sanger sequencing. The Kaplan-Meier curve assessed parasite clearance.
RESULTS: A total of 255 patients were enrolled in the study. The adequate clinical and parasitological response after PCR correction was 98.9% (95% CI 96.4-101.0%), 92.2% (85.0-98.5%) and 97.1% (93.1-101.0%) in Gaya, Tessaoua and Agadez, respectively. No adverse events were observed. Ten mutations (SNP) were found, including 7 synonyms (K248K, G690G, E691E, E612E, C469C, G496G, P718P) and 3 non-synonyms (N594K, R255K, V714S). Two mutations emerged: N594K and V714S. The R255K mutation detected in Southeast Asia was also detected. The pfdhpsK540E and pfdhfrI164L mutations associated with high levels of resistance are absent. There is a reversal of chloroquine resistance.
CONCLUSIONS: The study findings indicate that AL is effective and well tolerated for the treatment of uncomplicated malaria in three sites in Niger. The emergence of a pfk13 mutation requires additional testing such as the Ring Stage Assay and CRISPR/Cas9 to confirm the role of these emerging mutations. Trial registration NCT05070520, October 7, 2021.
METHODS: A multi-center, single-arm trial was conducted according to the 28-day World Health Organization (WHO) 2009 therapeutic efficacy study protocol. Children between 6 months and 15 years with confirmed uncomplicated Plasmodium falciparum infection and 1000-200,000 asexual parasites/μL of blood were enrolled and followed up for 28 days. Uncorrected and PCR-corrected efficacy results at day 28 were calculated, and molecular correction was performed by genotyping the msp1, msp2, and glurp genes. The pfk13, pfdhfr, pfdhps, pfcrt and pfmdr genes were analyzed by PCR and Sanger sequencing. The Kaplan-Meier curve assessed parasite clearance.
RESULTS: A total of 255 patients were enrolled in the study. The adequate clinical and parasitological response after PCR correction was 98.9% (95% CI 96.4-101.0%), 92.2% (85.0-98.5%) and 97.1% (93.1-101.0%) in Gaya, Tessaoua and Agadez, respectively. No adverse events were observed. Ten mutations (SNP) were found, including 7 synonyms (K248K, G690G, E691E, E612E, C469C, G496G, P718P) and 3 non-synonyms (N594K, R255K, V714S). Two mutations emerged: N594K and V714S. The R255K mutation detected in Southeast Asia was also detected. The pfdhpsK540E and pfdhfrI164L mutations associated with high levels of resistance are absent. There is a reversal of chloroquine resistance.
CONCLUSIONS: The study findings indicate that AL is effective and well tolerated for the treatment of uncomplicated malaria in three sites in Niger. The emergence of a pfk13 mutation requires additional testing such as the Ring Stage Assay and CRISPR/Cas9 to confirm the role of these emerging mutations. Trial registration NCT05070520, October 7, 2021.
摘要：
背景：监测治疗效果对于确保基于青蒿素的联合治疗（ACT）对疟疾的疗效很重要。尼日尔国家疟疾控制计划目前推荐的无并发症疟疾的一线治疗方法是蒿甲醚-lumefantrine（AL）。2020年，进行了一项体内研究，以评估三个前哨位点对AL的临床和寄生虫反应以及对药物的分子抗性：Agadez，Tessaoua和Gaya,在尼日尔。
方法：多中心，根据世界卫生组织(WHO)2009年28天疗效研究方案进行单组试验.纳入6个月至15岁的儿童，确诊为无并发症的恶性疟原虫感染和1000-200,000无性寄生虫/μL血液，并随访28天。计算第28天的未校正和PCR校正的功效结果，通过对msp1，msp2和glurp基因进行基因分型进行分子校正。pfk13，pfdhfr，pfdhps,通过PCR和Sanger测序分析pfcrt和pfmdr基因。Kaplan-Meier曲线评估寄生虫清除。
结果：本研究共纳入255例患者。PCR校正后的临床和寄生虫反应为98.9%（95%CI96.4-101.0%），加亚的92.2%(85.0-98.5%)和97.1%(93.1-101.0%)，Tessaoua和Agadez,分别。没有观察到不良事件。发现10个突变（SNP），包括7个同义词(K248K，G690G,E691E,E612E,C469C,G496G,P718P)和3个非同义词(N594K，R255K,V714S）。出现了两个突变：N594K和V714S。还检测到在东南亚检测到的R255K突变。不存在与高水平抗性相关的pfdhpsK540E和pfdhfrI164L突变。氯喹抗性存在逆转。
结论：研究结果表明，在尼日尔的三个地区，AL对治疗单纯性疟疾有效且耐受性良好。pfk13突变的出现需要额外的测试，如环阶段测定和CRISPR/Cas9，以确认这些新出现的突变的作用。试用注册NCT05070520，2021年10月7日。
方法：多中心，根据世界卫生组织(WHO)2009年28天疗效研究方案进行单组试验.纳入6个月至15岁的儿童，确诊为无并发症的恶性疟原虫感染和1000-200,000无性寄生虫/μL血液，并随访28天。计算第28天的未校正和PCR校正的功效结果，通过对msp1，msp2和glurp基因进行基因分型进行分子校正。pfk13，pfdhfr，pfdhps,通过PCR和Sanger测序分析pfcrt和pfmdr基因。Kaplan-Meier曲线评估寄生虫清除。
结果：本研究共纳入255例患者。PCR校正后的临床和寄生虫反应为98.9%（95%CI96.4-101.0%），加亚的92.2%(85.0-98.5%)和97.1%(93.1-101.0%)，Tessaoua和Agadez,分别。没有观察到不良事件。发现10个突变（SNP），包括7个同义词(K248K，G690G,E691E,E612E,C469C,G496G,P718P)和3个非同义词(N594K，R255K,V714S）。出现了两个突变：N594K和V714S。还检测到在东南亚检测到的R255K突变。不存在与高水平抗性相关的pfdhpsK540E和pfdhfrI164L突变。氯喹抗性存在逆转。
结论：研究结果表明，在尼日尔的三个地区，AL对治疗单纯性疟疾有效且耐受性良好。pfk13突变的出现需要额外的测试，如环阶段测定和CRISPR/Cas9，以确认这些新出现的突变的作用。试用注册NCT05070520，2021年10月7日。
