恶性疟原虫对哌喹敏感性的体外微量法的建立与应用

Eestablishment and application of in vitro microtechnique for assessment of sensitivity of Plasmodium falciparum to piperaquine

目的建立恶性疟原虫对哌喹敏感性的体外微量检测方法,以便在我国进行大规模的抗性调查。方法研制哌喹涂药板和供现场使用培养基,经实验室测试,后在海南省及云南省现场,比较体外微量法和体内四周法检测恶性疟原虫对哌喹敏感性的一致性。结果研制的哌喹涂药板及现场用的培养基质量稳定,4℃贮存哌喹涂药板和安瓿封装液体培养基有效期分别为14个月和2个月,哌喹涂药板判定敏感与抗性的临界药浓度为≥64pmol/井,体外微量法测得的结果与体内法一致。自1985年近30年现场监测,基本掌握我国恶性疟原虫对哌喹敏感性的状况,海南和云南两省恶性疟原虫对哌喹有高度抗性。结论成功建立起用体外微量法检测恶性疟原虫对哌喹的敏感性,为大规模抗药性检测提供了条件。

Objective To establishing an in vitro microtechnique for assessment of the sensitivity of Piasmodium flaciparum to piperaquine so as to carry out a large scale survey of drug resistance. Methods Piperaquine-coated plate and liquid medium were prepared and first tested in the laboratory, to verify its feasibility for assessment of the sensitivity of P. falciparum to piperaquine in vitro. After the piperaquine-eoated plate and medium were proved to be stable and reliable blood samples of clinical Plasmodium falcipaarum malaria cases were collected from Hainan and Yunnan and the sensitivity of P. falciparum to piperaquine were tested with in vivo 4-week test as a control. Results The piperaquine-coated plate and the medium for field application were effective and stable; the piperaquine-coated plate and the ampule-packaging of liquid medium stored at 4℃ could keep effective for 14 and 2 months respectively.The critical drug concentration of piperaquine- coated plate for assessing the resistance of Plasmodium falciparum was≥64pmol/well, showing that the result of in vitro microteehinque was consistent with in vivo 4-week test and could be used for assessing the sensitivity of P. falciparum to piperaquine after field application for nearly 30 years, the current status of sensitivity of P. falciparum to piperaquine in China were basically understood, indicating that P. falciparum in Hainan and Yunnan developed high resistance to piperaquine. Conclusion An in vitro microtechnique for assessment of sensitivity of P. falciparum to piper has been successively established, which has provided a favorable conditions for carrying out large scale assessment of drug resistance.