Novel phenotypic assays for the detection of artemisinin-resistant Plasmodium falciparum malaria in Cambodia: in-vitro and ex-vivo drug-response studies.
Novel phenotypic assays for the detection of artemisinin-resistant Plasmodium falciparum malaria in Cambodia: in-vitro and ex-vivo drug-response studies.
复制标题
用于检测柬埔寨抗青蒿素恶性疟原虫疟疾的新型表型测定：体外和离体药物反应研究。
DOI：
10.1016/s1473-3099(13)70252-4
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发表时间：
2013-12
影响因子：
56.3
通讯作者：
Menard, Didier
中科院分区：
文献类型：
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作者：
Witkowski, Benoit;Amaratunga, Chanaki;Khim, Nimol;Sreng, Sokunthea;Chim, Pheaktra;Kim, Saorin;Lim, Pharath;Mao, Sivanna;Sopha, Chantha;Sam, Baramey;Anderson, Jennifer M.;Duong, Socheat;Chuor, Char Meng;Taylor, Walter R. J.;Suon, Sella;Mercereau-Puijalon, Odile;Fairhurst, Rick M.;Menard, Didier
恶性疟原虫对青蒿素的耐药性延长了青蒿素单药治疗或以青蒿素为基础的联合治疗期间的寄生虫清除半衰期。缺乏体外和离体相关的青蒿素耐药性阻碍了这种表型的研究。我们的目的是评估体外环期生存试验（RSA）是否可以识别培养适应的恶性疟原虫分离物，这些分离物来自缓慢清除或快速清除感染的患者，研究体外环期生存试验中寄生虫对双氢青蒿素的分期敏感性，并评估体外环期生存试验是否可以识别耐青蒿素的恶性疟原虫感染。我们从2010年在柬埔寨Pursat进行的一项研究（在ClinicalTrials.gov注册，编号NCT00341003）中获得的具有较长和较短寄生虫清除半衰期的患者身上培养适应寄生虫，并使用新的体外生存试验来探索缓慢清除和快速清除寄生虫对双氢青蒿素的分期依赖性敏感性。2012年，我们在柬埔寨Pursat、柏威夏和Ratanakiri （NCT01736319）的前瞻性寄生虫清除研究中实施了RSA，以测量疟疾患者体内寄生虫的体外反应。采用Mann-Whitney U检验比较连续变量。用Spearman相关检验分析相关性。在0 - 3 h环期寄生虫中，13种慢清感染和13种快清感染的培养适应寄生虫体外存活率差异显著（10.88% vs 0.23%, p= 0.007）。离体存活率与体内寄生虫清除半衰期显著相关（n=30, r= 0.74, 95% CI 0.50 - 0.87; p< 0.0001）。0-3 h环期寄生虫的体外RSA为青蒿素耐药性的分子表征提供了一个平台。在需要监测青蒿素耐药性的地方，可以很容易地实施离体RSA。柬埔寨巴斯德研究所和校内研究项目，NIAID， NIH。
Artemisinin resistance in Plasmodium falciparum lengthens parasite clearance half-life during artemisinin monotherapy or artemisinin-based combination therapy. Absence of in-vitro and ex-vivo correlates of artemisinin resistance hinders study of this phenotype. We aimed to assess whether an in-vitro ring-stage survival assay (RSA) can identify culture-adapted P falciparum isolates from patients with slow-clearing or fast-clearing infections, to investigate the stage-dependent susceptibility of parasites to dihydroartemisinin in the in-vitro RSA, and to assess whether an ex-vivo RSA can identify artemisinin-resistant P falciparum infections. We culture-adapted parasites from patients with long and short parasite clearance half-lives from a study done in Pursat, Cambodia, in 2010 (registered with ClinicalTrials.gov, number NCT00341003) and used novel in-vitro survival assays to explore the stage-dependent susceptibility of slow-clearing and fast-clearing parasites to dihydroartemisinin. In 2012, we implemented the RSA in prospective parasite clearance studies in Pursat, Preah Vihear, and Ratanakiri, Cambodia (NCT01736319), to measure the ex-vivo responses of parasites from patients with malaria. Continuous variables were compared with the Mann-Whitney U test. Correlations were analysed with the Spearman correlation test. In-vitro survival rates of culture-adapted parasites from 13 slow-clearing and 13 fast-clearing infections differed significantly when assays were done on 0–3 h ring-stage parasites (10·88% vs 0·23%; p=0·007). Ex-vivo survival rates significantly correlated with in-vivo parasite clearance half-lives (n=30, r=0·74, 95% CI 0·50–0·87; p<0·0001). The in-vitro RSA of 0–3 h ring-stage parasites provides a platform for the molecular characterisation of artemisinin resistance. The ex-vivo RSA can be easily implemented where surveillance for artemisinin resistance is needed. Institut Pasteur du Cambodge and the Intramural Research Program, NIAID, NIH.