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目的了解海口市白纹伊蚊对溴氰菊酯和氯菊酯的抗药性,并检测其击倒抗性(kdr)基因突变型,阐明抗性表型与kdr基因突变的关系。方法在海口新埠岛、龙塘镇、石山镇、白沙门和花卉市场采集白纹伊蚊幼虫,应用浸渍法测定其对溴氰菊酯和氯菊酯杀虫剂的敏感性,以抗性倍数确定抗性级别。对应生物测定的样品扩增和分析白纹伊蚊的kdr基因部分片段,采用χ2检验比较抗性表型与kdr突变基因型的差异性。结果海口白纹伊蚊花卉市场和白沙门群体对溴氰菊酯和氯菊酯均产生抗性,花卉市场群体对溴氰菊酯的抗性倍数高达436.36,为高度抗性。龙塘镇和石山镇群体对氯菊酯均为敏感,对溴氰菊酯的抗性为低度(9.09)或中度(18.18)。新埠岛群体对溴氰菊酯和氯菊酯均为敏感。在对应检测的317只白纹伊蚊的kdr基因中,发现1 534位点存在点突变,检测到2种突变等位基因(TGC/C、TCC/S)。5种基因型和频率分别为野生型TTC/TTC(62.15%)、野生/突变型杂合子TTC/TGC(0.63%)和TTC/TCC(16.09%)、突变型纯合子TCC/TCC(20.82%)以及突变型杂合子TGC/TCC(0.32%)。χ2检验结果显示,接触菊酯类杀虫剂后敏感表型与抗性表型个体中,kdr基因型间的差异有统计学意义(P<0.05),在抗性表型群体中的kdr突变型基因型的频率(42.94%)大于敏感表型个体(31.97%)。结论海口市白纹伊蚊多个群体对溴氰菊酯和氯菊酯同时存在抗性,抗性表型与kdr突变基因型密切相关。本研究首次记录了kdr新的等位基因TCC/S。
Objective To understand the resistance of deltamethrin and permethrin to Aedes albopictus in Haikou city and to detect the kdr gene mutation and to elucidate the relationship between the resistance phenotype and kdr gene mutation. Methods Aedes albopictus larvae were collected from Xinbu Island, Longtang Town, Shishan Town, Baishamen and Flower Market in Haikou, and their susceptibility to deltamethrin and permethrin insecticide was determined by impregnation method. The resistance Multiple to determine resistance level. Corresponding biometrics samples were amplified and analyzed for part of the kdr gene of Aedes albopictus. Chi-square test was used to compare the differences between the resistant and kdr mutant genotypes. Results Both the Aedes albopictus flower market and Baishamen population in Haikou were resistant to both deltamethrin and permethrin. The flower market groups were highly resistant to deltamethrin up to 436.36. The communities of Longtang and Shishan were sensitive to permethrin and had low (9.09) or moderate (18.18) resistance to deltamethrin. The new port island population is sensitive to deltamethrin and permethrin. In the kdr gene of 317 Aedes albopictus detected, there were point mutations at 1 534 sites and two mutant alleles (TGC / C, TCC / S) were detected. The five genotypes and frequencies were wild type TTC / TTC (62.15%), wild / mutant heterozygotes TTC / TGC (0.63%) and TTC / TCC ) And mutant heterozygote TGC / TCC (0.32%). The results of χ2 test showed that there was significant difference in kdr genotypes between susceptible and resistant phenotypes after exposure to pyrethroid insecticides (P <0.05), kdr mutation in resistant phenotype population Genotype frequency (42.94%) than sensitive phenotype individuals (31.97%). CONCLUSIONS: Both populations of Aedes albopictus in Haikou are resistant to both deltamethrin and permethrin. The resistance phenotype is closely related to the kdr mutant genotype. This study first recorded kdr new allele TCC / S.