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目的监测云南省盈江县中缅边境恶性疟原虫对氯喹、哌喹的体外敏感性,了解两种抗疟药有无交叉耐药。方法选取2007~2009年当地病人体内的恶性疟原虫,体外培养成活共28株,在体外进行对氯喹、哌喹的半效抑虫浓度(IC_(50))值测定;并对疟原虫的Pfcrt基因片段扩增,测序,比对。结果 28株恶性疟原虫株体外氯喹的IC_(50)值为(463.9±256.0)nm;哌喹的IC_(50)值为(9.0±6.7nm)和野生株3D7相比,均有显著性差异(P<0.05)。28株(100%)恶性疟原虫对氯喹均耐药株;5/28(16.1%)的疟原虫对哌喹耐药;7/28(21.4%)的疟原虫对哌喹敏感。28株(100%)疟原虫的Pfcrt基因发现均有和氯喹耐药相关的76T变异。应用Pearson线性相关分析发现CQ和PPQ间存在叉抗药性(P<0.001)。结论云南省盈江县中缅边境的恶性疟原虫对氯喹、哌喹耐药的情况严峻,尤其是氯喹。
Objective To monitor the susceptibility of Plasmodium falciparum to chloroquine and piperaquine in Sino-Burma border in Yingjiang County, Yunnan Province, and to find out whether the two antimalarial drugs have cross-resistance. Methods Plasmodium falciparum was collected from local patients in 2007-2009, and 28 live strains were cultured in vitro. The half-inhibitory concentration (IC 50) of chloroquine and piperaquine was determined in vitro. The Pfcrt Gene fragment amplification, sequencing, alignment. Results The IC 50 value of chloroquine in vitro was (463.9 ± 256.0) nm in 28 strains of P. falciparum and that in piperacillin (9.0 ± 6.7 nm) was significantly higher than that of 3D7 (P <0.05). 28 (100%) P. falciparum isolates were resistant to chloroquine, 5/28 (16.1%) were resistant to pipefampin, and 7/28 (21.4%) were sensitive to pipefamponin. Pfcrt gene of 28 strains (100%) of Plasmodium were found to have 76T mutation associated with chloroquine resistance. Pearson linear correlation analysis revealed cross-resistance between CQ and PPQ (P <0.001). Conclusion Plasmodium falciparum in Sino-Burma border of Yingjiang County in Yunnan Province is very resistant to chloroquine and piperaquine, especially chloroquine.