摘要
目的检测上海市白纹伊蚊野外种群电压门控钠离子通道(VGSC)基因的基因型分布特点,了解白纹伊蚊击倒抗性(kdr)基因突变及趋势情况,为上海市蚊媒防制措施提供科学依据。方法于2024年9月—10月从上海市12个行政区13个地点采集白纹伊蚊幼虫/蛹,经实验室饲养羽化后,筛选健康雌蚊用于基因检测。采用离心柱法抽提基因组DNA,针对VGSC基因的DomainⅡ、Ⅲ、Ⅳ位点设计引物进行PCR扩增。扩增产物经琼脂糖凝胶电泳验证后送测,使用SeqMan软件比对序列峰图,确定突变位点及基因型。用Excel计算突变频率,并通过卡方检验比较区域间的差异。结果共检测455只白纹伊蚊,总体突变率为79.8%(363/455),13个群体均存在不同程度突变。V1016位点突变等位基因为GGA/G(5.0%,45/910),I1532位点突变等位基因为ACC/T(4.3%,39/910),F1534位点的2种突变等位基因分别为TCC/S(48.5%,441/910)和TGC/C(14.0%,127/910)。未发现3个位点同时突变的个体情况,两个位点同时突变的个体有52个(11.4%),V1016和F1534同时突变个体2个(0.4%),I1532和F1534位点同时突变个体50个(11.0%)。结论上海市白纹伊蚊群体KDR突变呈快速进化特征,其中1532位点(I/T)和1534位点(F/S)的组合突变型在多数监测区域稳定分布,需警惕协同抗性发展,且建立动态监测体系结合药剂轮换使用,构建多维度抗性治理策略,以延缓抗性基因扩散与累积。
Objective To investigate the genotype distribution of the voltage-gated sodium channel(VGSC)gene in wild Aedes albopictus field populations in Shanghai,characterize knockdown resistance(KDR)gene mutations and their trends,and provide scientific guidance for mosquito-borne disease control strategies in the region.Methods Larvae/pupae of Aedes albopictus were collected from 13 sites across 12 districts in Shanghai between September and October 2024.After being reared to adulthood in the laboratory,healthy female mosquitoes were selected for genetic analysis.Genomic DNA was extracted using a spin column method.Primers targeting DomainⅡ,Ⅲ,andⅣof the VGSC gene were designed for PCR amplification.Amplicons were verified by agarose gel electrophoresis,sequenced,and analyzed using SeqMan software to identify mutation sites and genotypes.Mutation frequencies were calculated using Excel,and regional differences were compared via chi-square tests(P<0.05 considered statistically significant).Results A total of 455 Aedes albopictus mosquitoes were analyzed,revealing an overall mutation rate of 79.8%(363/455),with all 13 populations exhibiting varying mutation frequencies.The frequencies of mutant alleles were 5.0%for V1016G(GGA/G,45/910),4.3%for I1532T(ACC/T,39/910),and for the F1534 locus, 48.5% for F1534S (TCC/S, 441/910) and 14.0% for F1534C (TGC/C, 127/910). No individuals carrying simultaneous mutations at all three loci were detected. However, 52 individuals (11.4%) harbored mutations at two loci: 2 individuals (0.4%) with concurrent mutations at V1016 and F1534, and 50 individuals (11.0%) with concurrent mutations at I1532 and F1534. Conclusion The Aedes albopictus population in Shanghai exhibits rapid KDR mutation evolution, with predominant combined mutations at loci I1532 (I/T) and F1534 (F/S) stably distributed across most surveillance regions. Vigilance is warranted regarding synergistic resistance development. A dynamic monitoring system, coupled with insecticide rotation and precision application, should be integrated into multi-dimensional resistance management strategies to delay resistance gene spread and accumulation.
作者
周秋明
盛望望
李佳璇
李翔宇
周琳瑛
许月萍
焦焱桐
袁浩
董昊炜
单文琪
彭恒
ZHOU Qiuming;SHENG Wangwang;LI Jiaxuan;LI Xiangyu;ZHOU Linying;XU Yueping;JIAO Yantong;YUAN Hao;DONG Haowei;SHAN Wenqi;PENG Heng(Department of Naval Environmental and Occupational Health,Faculty of Naval Medicine,Naval Medical University,Shanghai 200433,China;Integrated Department,The 72nd Group Army Hospital of PLA,Huzhou,Zhejiang 313000,China;The Fifth Brigade of Undergraduates,College of Basic Medicine Sciences,Naval Medical University,Shanghai 200433,China;Department of Medical Microbiology and Parasitology,Naval Medical University,Shanghai 200433,China;Department of Casualty Management,The 72nd Group Army Hospital of PLA,Huzhou,Zhejiang 313000,China;Department of Neuroendocrinology,The 72nd Group Army Hospital of PLA,Huzhou,Zhejiang 313000,China)
出处
《中国热带医学》
北大核心
2025年第11期1446-1450,1498,共6页
China Tropical Medicine
基金
上海市卫生健康委员会卫生行业临床研究专项(No.202240144)。
关键词
白纹伊蚊
击倒抗性基因
基因突变
抗药性
蚊媒防制
Aedes albopictus
knockdown resistance gene
gene mutation
insecticide resistance
mosquito vector control
