逯小强负责实验设计、实验操作、数据分析、论文撰写，杨滨瑜和赵琼瑶负责样品采集和实验操作，刘艳春和孔新明负责实验准备，贾永朝负责结果核验和论文修改。
收稿日期: 2025-09-03
修回日期: 2025-11-10
网络出版日期: 2026-05-21
基金资助
广元市社会发展领域重点研发项目(21ZDYF0115)
Frequency of I1532T mutations in the voltage-gated sodium channel of Aedes albopictus in Guangyuan City
Received date: 2025-09-03
Revised date: 2025-11-10
Online published: 2026-05-21
Supported by
Key Research and Development Project in the Field of Social Development of Guangyuan City(21ZDYF0115)
为了解四川省广元市白纹伊蚊电压门控钠离子通道（vgsc）基因I1532T突变频率，于2024年6—7月在广元市7个县（区）居民区外以CO₂灯诱法采集白纹伊蚊成蚊，提取基因组DNA，聚合酶链式反应-限制性片段长度多态性（PCR-RFLP）检测白纹伊蚊vgsc 1532位点的基因型。通过测序样品验证PCR-RFLP技术的检测效果。使用Genepop 4.7进行种群哈代温伯格平衡（HWE）检验。使用SPSS 25.0对各县（区）间等位基因频率进行Fisher精确检验。共检测白纹伊蚊208只。PCR结果显示，所有样品均扩增出约220 bp的条带，与预期大小相符。广元市vgsc野生型1532I/I纯合子、野生型/突变型1532I/T杂合子、抗性1532T/T纯合子频率分别为87.98%（183/208）、10.58%（22/208）和1.44%（3/208）。7个县（区）中，剑阁县、利州区、青川县、旺苍县、昭化区野生型/突变型1532I/T杂合子频率分别为12.50%（4/32）、9.38%（3/32）、17.86%（5/28）、28.13%（9/32）、1/16，朝天区和苍溪县未发现；利州区和旺苍县抗性1532T/T纯合子频率分别为6.25%（2/32）和3.13%（1/32），其他县（区）未发现。广元市vgsc野生型1532I和突变型1532T等位基因的频率分别为93.27%（388/416）、6.73%（28/416）。朝天区、苍溪县、剑阁县、利州区、青川县、旺苍县和昭化区突变型1532T等位基因频率分别为0（0/64）、0（0/72）、6.25%（4/64）、10.94%（7/64）、8.93%（5/56）、17.19%（11/64）、3.13%（1/32），差异有统计学意义（P < 0.05）。HWE检验显示，利州区种群显著偏离遗传平衡（P < 0.05），剑阁县、青川县和旺苍县种群均符合遗传平衡（P > 0.05），其余3个县（区）因未发现1532T突变（朝天区和苍溪县）或突变频率低（昭化区），不适合做HWE检验。验证结果显示，PCR-RFLP与Sanger测序结果符合率100%。本研究发现广元市白纹伊蚊种群vgsc I1532T突变频率较高，并在利州区和旺苍县检测到抗性纯合子个体。
关键词： 白纹伊蚊; 聚合酶链式反应-限制性片段长度多态性; 电压门控钠离子通道; 击倒抗性突变
逯小强 , 杨滨瑜 , 赵琼瑶 , 孔新明 , 刘艳春 , 贾永朝 . 广元市白纹伊蚊电压门控钠离子通道I1532T突变频率调查分析[J]. 中国寄生虫学与寄生虫病杂志, 2026 , 44(3) : 443 -447 . DOI: 10.12140/j.issn.1000-7423.2026.03.019
To investigate the frequency of the I1532T mutation in the voltage-gated sodium channel (vgsc) of Aedes albopictus in Guangyuan City, Sichuan Province, adult Ae. albopictus mosquitoes were collected using CO₂-baited light trapsfrom residential areas in seven counties (districts) of Guangyuan City between June and July in 2024. Genomic DNA was extracted from mosquitoes, and the genotype at the vgsc 1532 locus was detected using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. The detection performance of PCR-RFLP assay was validated by sequencing. Hardy-Weinberg equilibrium (HWE) test for populations was assessed using the software Genepop 4.7, and Fisher’s exact test was used to compare allele frequencies across counties (districts) using the software SPSS 25. A total of 208 Ae. albopictus mosquitoes were detected. PCR amplification of all samples showed bands with approximately 220 bp in length, which was consistent with the expected size. The frequencies of the vgsc wild-type homozygous 1532I/I genotype, wild-type/mutant heterozygous 1532I/T genotype, and resistant homozygous 1532T/T genotype were 87.98% (183/208), 10.58% (22/208), and 1.44% (3/208) in Guangyuan City, respectively. Among 7 counties (districts), the frequencies of wild-type/mutant heterozygous 1532I/T genotype were 12.5% (4/32), 9.38% (3/32), 17.86% (5/28), 28.13% (9/32) and 1/16 in Jiange County, Lizhou District, Qingchuan County, Wangcang County and Zhaohua District, respectively, while no heterozygous genotype were found in Chaotian District or Cangxi County. The frequencies of resistant homozygous 1532T/T genotype were 6.25% (2/32) and 3.13% (1/32) in Lizhou District and Wangcang County, respectively, and this homozygous genotype were not detected in other counties (districts). The frequencies of the vgsc wild-type 1532I and mutant 1532T alleles were 93.27% (388/416) and 6.73% (28/416) in Guangyuan City, respectively, and the frequencies of mutant 1532T alleles were 0 (0/64), 0 (0/72), 6.25% (4/64), 10.94% (7/64), 8.93% (5/56), 17.19% (11/64), and 3.13% (1/32) in Chaotian District, Cangxi County, Jiange County, Lizhou District, Qingchuan County, Wangcang County, and Zhaohua District, respectively (P < 0.05). HWE test showed that the Ae. albopictus population in Lizhou District deviated significantly from equilibrium (P < 0.05), and Ae. albopictus populations from Jiange County, Qingchuan County, and Wangcang County were at equilibrium (P > 0.05), while Ae. albopictus populations from other three counties (districts) were not suitable for HWE tests due to absence of the 1532T mutation (Chaotian District and Cangxi County) or low mutation frequency (Zhaohua District). The verification results showed that the coincidence rate between PCR-RFLP and sanger sequencing results was 100%. This study indicates a relatively high frequency of the vgsc I1532T mutation in Ae. albopictus populations from Guangyuan City, with resistant homozygotes detected in Lizhou District and Wangcang County.
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