云南省中华按蚊电压门控钠离子通道kdr突变频率调查  

Investigation of the frequency of kdr mutation of voltage-gated sodium channel in Anopheles sinensis in Yunnan Province

作　　者：曾旭灿[1] 林祖锐[1] 许翔 吴超[1] 魏春[1] 孙晓东[1] 杨锐[1] ZENG Xu-can;LIN Zu-rui;XU Xiang;WU Chao;WEI Chun;SUN Xiao-dong;YANG Rui(Yunnan Provincial Key Laboratory of Vector-borne Diseases Control and Research&Yunnan Innovative Team of Key Techniques for Vector-borne Disease Control and Prevention of Yunnan Institute of Parasitic Diseases,Pu'er,Yunnan 665000,China)

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

出　　处：《中国热带医学》2022年第10期904-908,共5页China Tropical Medicine

基　　金：国家自然科学基金项目(No.81960374);云南省重点研发计划项目(No.202103AQ100001)。

摘　　要：目的掌握云南省中华按蚊电压门控钠离子通道击倒抗性(knockdown resistance,kdr)基因突变情况。方法2018—2019年,在云南省罗平县、绥江县、腾冲市、盈江县、元江县和勐腊县采集蚊虫。采集的蚊虫经形态学鉴定为中华按蚊,用试剂盒提取成蚊基因组DNA。DNA模板经PCR扩增后测序,测序结果经NCBI同源性比对确定为中华按蚊。扩增中华按蚊钠离子通道ⅡS5、ⅡS6片段后测序,测序结果用DNAMAN软件进行多序列比对,用BioEdit软件对测序峰图逐条分析突变情况,确定kdr等位基因类型和基因型并计算频率。结果本次调查扩增得到287条序列,测序峰图显示1014位点共有3种等位基因,包括野生型TTG/L(89.20%)和突变型TTT/F(9.76%)、TCG/S(1.04%);5种基因型:野生型纯合子L/L(85.02%),突变型纯合子F/F(6.27%)、S/S(0.35%),突变型杂合子L/F(6.97%)和L/S(1.39%)。在6个采集点中除绥江县外,均以野生型等位基因TTG/L为主。腾冲市野生型等位基因频率最高(100.00%),即未检测到突变,而其余各县1014位点均出现不同程度突变。绥江县突变型等位基因频率最高,达到55.68%。5种基因型中,罗平县、勐腊县和绥江县各有2种突变型,盈江县和元江县有1种突变型杂合子L/F。结论云南省大部分地区中华按蚊kdr基因仍以野生型L1014(TTG/L)为主,kdr突变类型主要为L1014F,其次是L1014S,且突变频率低于中国中部省份。Objective To understand the kdr(knockdown resistance,kdr)gene mutation of the voltage-gated sodium channel(VGSC)of Anopheles sinensis in Yunnan Province.Methods From 2018 to 2019,mosquitoes were collected in Luoping County,Suijiang County,Tengchong City,Yingjiang County,Yuanjiang County and Mengla County in Yunnan Province.The collected mosquitoes were morphologically identified as Anopheles sinensis and genomic DNA was extracted by kits.The DNA templates were sequenced after PCR amplification and the sequencing results were identify as Anopheles sinensis by homology alignment in NCBI.After theⅡS5 andⅡS6 fragments of the sodium channels in Anopheles sinensis were amplified and sequenced,the sequencing results were multiple aligned by DNAMAN software,and the mutations were analyzed one by one with BioEdit software to determine the kdr allele types and genotypes,and the frequencies were calculated.Results This survey amplified 287 sequences,and the sequence maps showed that 1014 loci had three alleles,including wild type TTG/L(89.20%),mutant type TTT/F(9.76%)and TCG/S(1.04%).Five genotypes:homozygous wildtype L/L(85.02%),homozygous mutant F/F(6.27%)and S/S(0.35%),heterozygous mutant L/F(6.97%)and L/S(1.39%).The wild type allele TTG/L was the main allele in six sampling sites except Suijiang County.The frequency of wild type allele in Tengchong City was the highest(100.00%).That is,no mutation was detected,while the rest of counties occurred different degrees of mutation at 1014 loci.The frequency of mutant allele in Suijiang County was the highest,reaching 55.68%.Luoping County,Mengla County and Suijiang County had two mutant types.Yingjiang County and Yuanjiang County had one heterozygous mutant L/F.Conclusion Wild type L1014(TTG/L)is still dominant in most areas of Yunnan Province.The kdr mutation type is mainly L1014F,followed by L1014S,and the mutation frequency is lower than that in central provinces of China.

关 键 词：中华按蚊 击倒抗性 钠离子通道 云南 

分 类 号：R531.3[医药卫生—内科学]