中缅边境地区恶性疟原虫氯喹抗性相关基因的多态性分析  

Polymorphisms of Plasmodium falciparum chloroquine resistant genes in China-Myanmar border

作　　者：叶升玉 李春富[1] 唐烨榕 叶润 周红宁[1] YE Sheng-yu;LI Chung-fu;TANG Ye-rong;YE Run;ZHOU Hong-ning(The Yunnan Provincial Key Laboratory of Insect-borne Infectious Diseases Control&the Yunnan Provincial Key Technology Innovation Team for Prevention and Control of Insect Vectors of Yunnan Institute of Parasitic Diseases,Pu'er Yunnan 665000,China;Department of Tropical Diseases,Navy Medical University)

机构地区：[1]云南省寄生虫病防治所云南省虫媒传染病防控研究重点实验室云南省虫媒传染病防控关键技术创新团队,云南普洱665000 [2]中国人民解放军海军军医大学热带病学教研室

出　　处：《中国病原生物学杂志》2022年第6期724-729,共6页Journal of Pathogen Biology

基　　金：云南省重点研发计划项目(No.202103AQ100001);澜湄合作专项基金项目(No.2020399)。

摘　　要：目的 分析中缅边境地区恶性疟原虫氯喹抗性相关基因的多态性。方法 收集2001-2012年中国云南省西双版纳州景洪市、缅甸克钦邦拉咱市和掸邦第二特区勐冒县恶性疟病例滤纸血标本175份,采用巢式PCR法扩增恶性疟原虫氯喹抗性转运蛋白基因(Plasmodium falciparum chloroquine resistance transporter gene,pfcrt)基因第72-76位点片段和多药耐药基因1(Plasmodium falciparum multidrug resistance gene 1,pfmdr1)基因第86-1246位点片段,并对扩增产物测序分析。结果 共检测到pfcrt基因5种突变型(CVIEK,CVIDK,SVMNT,CVIET,CVIDT),该3个地区均以三重突变型(CVIET)为主,其中,中国云南西双版纳CVIET由2001年的76.7%上升至2006年的100%,缅甸拉咱市由2001年的75%上升至2007年的95%,但2012年又下降至77.5%,缅甸勐冒县2009年该突变型占比为67.5%。共检测到pfmdr1基因5种单突变型(N86Y,E130K,Y184F,S1034C,F1226Y)和2种双重突变型(N86Y/F1226Y,Y184F/F1226Y),其中2001年西双版纳pfmdr1基因野生型占36.7%,N86Y和Y184F突变率均为20%,F1226Y突变率为16.7%,而2006年以F1226Y突变为主占40%,且检测到二重突变型(N86Y/F1226Y和Y184F/F1226Y)占8%;缅甸拉咱市pfmdr1基因Y184F突变率从2001年的25%升高至2007年的35%和2012年的37.5%,该3年中野生型分别占65%、40%和55%;2009年缅甸勐冒县pfmdr1基因野生型比例为47.5%,Y184和N86Y单突变型分别为27.5%和25%。结论 中缅边境恶性疟原虫pfcrt基因均以三重突变型(CVIET)为主,氯喹抗性均较高,可为中缅边境地区恶性疟的防治提供参考。Objective To monitoranalyze the polymorphisms of Plasmodium falciparum chloroquine resistant genes in three areas along the China Myanmar border.the dynamic changes of chloroquine resistance of Plasmodium falciparum in three areas along the China Myanmar border by detecting the mutations of Plasmodium falciparum chloroquine resistance transporter gene(pfcrt) and Plasmodium falciparum multidrug resistance gene 1(pfmdr1) gene.Methods Totally 175 P.falciparum filter paper blood samples were collected in Jinghong City,Xishuangbanna Prefecture of Yunnan Province,China and Lazan City of Kachin State and Mengmao County,the Second Special Zone of Shan State in Myanmar from 2001 to 2012.The fragments of Plasmodium falciparum chloroquine resistance transporter gene(pfcrt)pfcrt gene(positions 72 to 76) and Plasmodium falciparum multidrug resistance gene 1(pfmdr1) pfmdr1 gene(positions 86 to 184,and positions 1034 to 1246) were amplified by Nested PCR,and then products were sequenced and statistically analyzed.Results Five haplotypes(CVIEK,CVIDK,SVMNT,CVIET and CVIDT) were detected from positions 72 to 76 of pfcrt gene,of which the triple mutants(CVIET) was the major haplotype in three areas.The percentage of haplotype CVIET was 76.7% in 2001 and increased to 100% in 2006 in Xishuangbanna.In Lazan City,the percentage of haplotype CVIET increased from 75% in 2001 to 95% in 2007 but decreased to 77.5% in 2012,and it accounted for 67.5% in Mengmao County in 2009.Single mutants(N86 Y,E130 K,Y184 F,S1034 C and F1226 Y) and double mutants(N86 Y/F1226 Y and Y184 F/F1226 Y) were detected in the pfmdr1 gene.In Xishuangbanna,isolates with wild type pfmdr1 gene accounted for 36.7%,the proportions of the mutant N86 Y,Y184 F and F1226 Y was 20%,20% and 16.7% in 2001,respectively.While in 2006,the rate of mutant F1226 Y increased to 40%,and double mutants(N86 Y/F1226 Y and Y184 F/F1226 Y) was also identified,which accounted for 8% in total.In Lazan City,the rate of single mutants Y184 F of pfmdr1 increased from 25% in 2001 to 35% in 2007

关 键 词：恶性疟原虫 pfcrt基因 pfmdr1基因 多态性 中缅边境地区 

分 类 号：R382.31[医药卫生—医学寄生虫学]