云南省抗氯喹恶性疟原虫双氢青蒿素抗性株体外培育与单克隆品系的建立  

Creation of cloned strains of chloroquine-resistant Plasmodium falciparumfrom Yunnan,China with resistance to dihydroartemisinin

作　　者：杨亚明[1] 李雪平[2] 张苍林[1] 李奔福[1] 刘慧[1] 李丽[1] 

机构地区：[1]云南省寄生虫病防治所,云南普洱665000 [2]大理学院病原与媒介生物研究所

出　　处：《中国病原生物学杂志》2013年第8期714-717,共4页Journal of Pathogen Biology

基　　金：云南省应用基础研究项目(No.2010CD132)

摘　　要：目的建立云南省恶性疟原虫抗双氢青蒿素的单克隆品系,为开展恶性疟原虫对青蒿素类的抗药性研究奠定基础。方法体外长期连续培养恶性疟原虫,采用体外间断双氢青蒿素药物刺激法对恶性疟原虫株进行抗药性培育,待达到一定抗药性后采用有限稀释法对虫株进行克隆,采用体外微量法测定克隆虫株对双氢青蒿素、青蒿素、氯喹的敏感性,并对其生物学特性进行鉴定。结果获得2个云南氯喹抗性恶性疟原虫对双氢青蒿素抗性的单克隆品系C10和H6,IC50值分别为39.01nmol/L和26.91nmol/L。结论经过8次体外间断药物刺激和筛选,获得2株恶性疟双氢青蒿素抗性单克隆品系,其药物耐受性水平与现场抗性株接近。Objectives To establish monoclonal strains of Plasmodium falciparumfrom Yunnan Province with resistance to dihydroartemisinin and to lay the foundation for study of the resistance of P.falciparumto artemisinin-based drugs.Methods Strains of P.falciparum were cultured via long-term continuous in vitro culturing.Strains were intermittently challenged with dihydroartemisinin in vitro.After the strains developed a certain level of resistance to dihydroartemisinin according to a microtest assay,they were cloned using a limited dilution.Biological characteristics of sensitivity to dihydroartemisinin,artemisinin,and chloroquine in vitro were identified.Results Two monoclonal strains of dihydroartemisinin-resistant P.falciparum were obtained from chloroquine resistant P.falciparumoriginating in Yunnan.The IC50of the C10cloned strain was 39.01nmol/L and the IC50 of the H6 cloned strain was 26.91nmol/ L.Conclusion Two monoclonal strains of dihydroartemisinin-resistant P.falciparum were obtained after the parasites were intermittently challenged with 8rounds of dihydroartemisinin in vitro.These strains had a level of drug resistance close to that of resistant strains from Yunnan.

关 键 词：疟原虫 恶性 双氢青蒿素 抗药性 克隆株 

分 类 号：R382.31[医药卫生—医学寄生虫学]