中国恶性疟原虫氯喹抗药性相关基因Pfcrt 72-76位点基因型分析  

The polymorphisms of Pfcrt 72-76 in Plasmodium falciparum isolated from China and their relationships with chloroquine resistance

作　　者：张国庆[1] 官亚宜[1] 胡铃[1] 汤林华[1] 冯晓萍[1] 蔡玥[1] 姚俊敏[1] 刘德全[1] 

机构地区：[1]中国疾病预防控制中心寄生虫病预防控制所,卫生部寄生虫病原与媒介生物学重点实验室,世界卫生组织疟疾,血吸虫病和丝虫病合作中心,上海200025

出　　处：《中国人兽共患病学报》2009年第8期725-728,共4页Chinese Journal of Zoonoses

基　　金：日本国立传染病研究所资助中国疾病预防控制中心青年科研基金课题(2009A105)

摘　　要：目的了解我国云南和海南恶性疟原虫分离株Pfcrt基因72-76位点基因型,并分析该位置不同基因型与恶性疟原虫对氯喹敏感性的关系。方法从我国恶性疟流行区云南和海南省采集恶性疟现症患者血样,根据恶性疟原虫Pfcrt基因序列设计引物,进行巢式PCR扩增和测序。采用WHO制定的体外微量法测定同批血样中恶性疟原虫对氯喹的敏感性。结果云南、海南省恶性疟原虫Pfcrt基因第72-76位编码的氨基酸单元型在氯喹敏感株均为CVMNK。其中海南的氯喹抗性株Pfcrt基因第72-76位编码的氨基酸为单一的CVIET单元型,云南氯喹抗性株中除一株为CVIKT,另1株为SVMNT外,均为CVIET单元型。结论我国云南、海南省恶性疟原虫氯喹敏感株Pfcrt基因第72-76位编码的氨基酸基因型为CVMNK。抗性株主要为CVIET型,Pfcrt基因第72-76位编码氨基酸基因型可作为氯喹抗性分子标志。To investigate the polymorphisms of Pfcrt 72- 76 in Plasmodium falciparum isolated from Hainan and Yunnan provinces,so as to determine the correlation between the prevalence of polymorphisms and the level of chloroquine re- sistance of P. falciparum isolates. Blood samples were taken from the cases detected as falciparum malaria in Yunnan and Hainan provinces. Nested PCR was applied to amplify the fragments of Pfcrt gene including the 72^nd to 76^th condon. DNA sequencing was carried out for the nested PCR products. Chloroquine resistance of the same isolates was measured by the in vitro microtest. The chloroquine sensitivity of P. falciparurn isolates from Yunnan and Hainan carried CVMNK genotype in Pfcrt 72--76, And the chloroquine resistant P. falciparum isolates from Hainan carried CVIET genotype in Pfcrt 72--76. For the chloroquine resistant isolates from Yunnan, one carried carry CVIKT genotype, one carried SVMNT genotype, and the others carried CVIET genotype. The chloroquine sensitivity P. falciparurn isolates from China carry CVMNK genotype in Pfcrt 72-- 76; most of the chloroquine resistant isolates CVIET genotype in Pfcrt 72--76; The polymorphisms in Pfcrt 72 76 may be used as a molecular marker in the surveillance of the changes of chloroquine resistance in P. falciparurn.

关 键 词：恶性疟原虫 药物抗性 单核苷酸多态性 氯喹 

分 类 号：R382[医药卫生—医学寄生虫学]