深圳市2022—2023年输入性恶性疟原虫耐药基因突变情况  

Drug-resistant gene mutations in imported Plasmodium falciparum in Shenzhen,2022-2023

作　　者：刘小莲 高世同[1] 李媛[1] 唐屹君[1] 张倩[1] 彭博[1] 阳帆[1] 张仁利[1] 黄达娜[1] LIU Xiaolian;GAO Shitong;LI Yuan;TANG Yijun;ZHANG Qian;PENG Bo;YANG Fan;ZHANG Renli;HUANG Dana(Shenzhen Center for Disease Control and Prevention,Shenzhen,Guangdong 518055,China)

机构地区：[1]深圳市疾病预防控制中心,广东深圳518055

出　　处：《中国热带医学》2024年第4期382-387,共6页China Tropical Medicine

基　　金：深圳医疗卫生“三名工程”项目(No.SZSM202211023)。

摘　　要：目的分析了解深圳市输入性恶性疟原虫耐药基因突变情况,帮助评估抗疟药的疗效和指导有效用药。方法收集2022—2023年从境外输入深圳市的恶性疟荧光定量聚合酶链反应(polymerase chain reaction,PCR)阳性的样本85例,提取基因组DNA,使用巢式PCR扩增耐药基因:恶性疟原虫Kelch螺旋体蛋白基因(Plasmodium falciparum Kelch 13,PfK13)、多药耐药基因1(multidrug resistance 1,Pfmdr1)、氯喹抗性转运基因(chloroquine resistance transporter,Pfcrt)、二氢叶酸还原酶基因(dihydrofolate reductase,Pfdhfr)和二氢蝶酸合成酶基因(dihydropteroate synthase,Pfdhps),并进行双向测序,使用MEGA11.06软件分析这些耐药基因突变情况。结果研究发现PfK13错义突变(S549P)1例,同义突变4例。Pfmdr162.69%样本存在Y184F突变,未检测到N86Y突变体。Pfcrt基因中,未检测到72和73位点的突变,M74I、N75E和K76T突变分别占17.46%、15.87%和15.87%。Pfcrt基因野生型占主导(82.54%,52例),其次是I_(74)E_(75)T_(76)三重突变体(15.87%,10例)。Pfdhfr最常见的突变型为I_(51)R_(59)N_(108)(91.78%,67例),其次是野生型(2.74%,2例)。Pfdhps野生型占60.32%(38例),单突变K540E为最常见突变类型,检测到S436A、G437A、K540E、A581G、A613S、I431V、G556K、G579E位点突变。Pfdhfr-Pfdhps组合突变中,I_(51)R_(59)N_(108)-E540是频率最高的组合突变(11.48%),59.02%样本为Pfdhfr单独突变体。结论在本研究中,PfK13突变率低,且无报导的耐药突变,Y184F成为Pfmdr1突变主导,未发现有N86Y。Pfcrt野生型为主,其次为I_(74)E_(75)T_(76)三重突变体,Pfdhfr I_(51)R_(59)N_(108)三重突变非常普遍,本研究并未发现PfdhfrPfdhps完全抗性和超抗性突变体,但有其他的五重七重突变型。在未来的工作中,需要继续加强疟原虫耐药基因的监测,同时进一步结合体内疗效的监测来有效指导临床用药。Objective To analyze and understand the mutations of drug resistance genes in imported Plasmodium falciparum in Shenzhen,aiming to assess the efficacy of antimalarial drugs and guide effective drug use.Methods A total of 85 samples from individuals with imported Plasmodium falciparum confirmed by fluorescence quantitative polymerase chain reaction(PCR)in Shenzhen from 2022 to 2023 were collected and genomic DNA was extracted.Nested PCR was used to amplify resistance genes,including Plasmodium falciparum Kelch 13(PfK13),multidrug resistance gene 1(Pfmdr1),chloroquine resistance transporter(Pfcrt),dihydrofolate reductase(Pfdhfr),and dihydropteroate synthase(Pfdhps)genes.Bidirectional sequencing was conducted,and mutations in these resistance genes were analyzed using MEGA11.06 software.Results The study found one missense mutation(S549P)and four synonymous mutations in PfK13.For Pfmdr1,62.69%of the samples showed Y184F mutation,and no N86Y mutation was detected.No mutations at positions 72 and 73 were detected in the Pfcrt gene,while mutations at M74I,N75E,and K76T accounted for 17.46%,15.87%,and 15.87%,respectively.The wild-type of Pfcrt gene is dominant(82.54%,52),followed by the triple mutant I_(74)E_(75)T_(76)(15.87%,10).The most common mutation type for Pfdhfr is I_(51)R_(59)N_(108)(91.78%,67),followed by the wild type(2.74%,2).More than half(60.32%,38)of the Pfdhps samples were wild-type,with single mutation K540E being the most common mutation type.S436A,G437A,K540E,A581G,A613S,I431V,G556K,and G579E site mutations were detected.Among the Pfdhfr-Pfdhps combination mutations,I_(51)R_(59)N_(108)-E540 was the most frequent combination mutation(11.48%),with 59.02%of samples showing solitary Pfdhfr mutations.Conclusions In this study,PfK13 mutation rates were low,with no reported resistance mutations.The Y184F mutation emerged as the dominant Pfmdr1 mutation,with no detection of N86Y.For Pfcrt,the wild-type was dominant,followed by the I_(74)E_(75)T_(76)triple mutation variant.Triple mutant I_(51)R_(59)N_(108)of P

关 键 词：恶性疟原虫 输入性 耐药性 基因 突变 

分 类 号：R531[医药卫生—内科学]