境外输入性恶性疟原虫药物抗性基因多态性分析  

Analysis of polymorphisms in anti-malarial resistance genes in imported Plasmodium falciparum

作　　者：杨成运[1] 李素华[1] 张雅兰[1] 周瑞敏[1] 刘颖[1] 钱丹[1] 赵玉玲[1] 许汴利[1] 张红卫[1] YANG Cheng-yun;LI Su-hua;ZHANG Ya lan;ZHOU Rui-min;LIU Ying;QIAN Dan;ZHAO Yu-ling;XU Bian-li;ZHANG Hong-wei(Henan Center for Disease Control and Prevention,Henan Key Laboratory of Pathogenic Organisms,Zhengzhou 450016,China)

机构地区：[1]河南省疾病预防控制中心河南省传染病病原生物重点实验室,河南郑州450016

出　　处：《中国病原生物学杂志》2018年第9期998-1001,共4页Journal of Pathogen Biology

基　　金：河南省医学科技攻关计划项目(No.201702274)

摘　　要：目的了解河南省输入性恶性疟原虫相关抗性基因的突变情况。方法采集河南省2016年自非洲劳务返乡的131例输入性恶性疟患者血样,提取DNA,采用Pfdhfr、Pfmdr1和K13基因序列引物进行巢式PCR扩增并测序,对测序结果进行序列比对,分析基因突变情况。结果 131例输入性恶性疟患者自19个非洲国家务工返乡。131份血样均扩增出Pfdhfr、Pfmdr1和K13基因片段。其中Pfdhfr基因51,59和108位点氨基酸双突变NRNI型占2.29%,ICNI型占10.69%,三突变IRNI型占85.49%,野生型占1.53%,未见四突变。Pfmdr1基因86突变率分别为9.16%,K13突变率为11.45%,共15份血样检测到9个位点突变。结论 2016年河南省输入性恶性疟原虫Pfdhfr、Pfmdr1和K13基因均有不同程度的突变,其中Pfdhfr基因三突变型所占比例较高,K13基因未检测到与青蒿素抗性相关的突变。Objectives To ascertain mutations in genes associated with drug resistance in imported P.falciparumin Henan Province. Methods Blood samples and information were collected from 131 migrant workers with P.falciparum malaria who returned to Henan Province from Africa in 2016.Genomic DNA was extracted,and Pfdhfr,Pfmdr1,and K13 were amplified with nested PCR using specific primers.The PCR products were sequenced.Sequencing was analyzed using BLAST,and sequences were aligned with reference sequences using BioEdit Sequence Alignment Editor.Polymorphisms in and the distribution of Pfdhfr,Pfmdr1,and K13 were analyzed. Results The 131 cases of imported malaria were from 19 African countries.Pfdhfr,Pfmdr1,and K13 were amplified from all of the blood samples.Sequencing revealed that there were various mutations at position 51,59,and 108 in the Pfdhfr gene,including double mutations and triple mutations.The double mutations NRNI and ICNI were respectively noted at a rate of 2.29% and 10.69%,respectively.The triple mutation IRNI was noted at a rate of 85.49%.Wild-type NCSI was noted at a rate of 1.53%,and no quadruple mutations were detected.P.falciparuminfecting the 131 patients had 12 mutations at position 86,and Pfmdr1 was noted at a rate of 9.16%.Twelve patients returned from 6 countries.K13 was noted at a rate of 11.45%.Nine point mutations were noted in 15 of the 131 isolates.The C580 Y mutation plays a predominant role in resistance to artemisinin but was not detected in the samples.Fifteen patients returned from 6 countries：Angola,Congo,Ghana,Nigeria,Uganda,and Cameroon. Conclusion There were mutations in Pfdhfr,Pfmdr1,and K13 in imported P.falciparumin Henan Province.The triple mutation Pfdhfr was found most often.No mutations in the K13 gene that were associated with artemisinin resistance were noted.

关 键 词：输入性恶性疟 Pfdhfr基因 Pfmdr1基因 K13基因 突变 河南省 

分 类 号：R382.31[医药卫生—医学寄生虫学]