海口市2018年白纹伊蚊击倒抗性基因型分布研究
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摘要
目的了解海南省海口市登革热媒介白纹伊蚊的击倒抗性基因(kdr)在不同区域的基因型及其分布特点,并分析其抗药性产生的影响因素,为白纹伊蚊化学防控提供依据。方法 2018年6-7月在海口市东、南、西、北、中5个方位,采集白纹伊蚊幼虫及蛹,饲养至成蚊,经形态学方法鉴定后,用无水乙醇溶液浸泡,并于-20℃冻存备用。提取单只蚊虫的DNA,PCR扩增神经细胞膜上电压门控钠离子通道(VGSC)基因部分片段,测序后分析kdr基因突变情况。结果对海口市不同方位采集的94只白纹伊蚊的检测结果显示,其kdr基因F1534位点存在突变。1534位点共有4种等位基因,即野生型TTC/F(113/60.11%),突变型TCC/S(66/35.11%)、TCG/S(1/0.53%)和TGC/C(8/4.26%);有6种基因型,分别为野生型纯合子F/F(39/41.49%),野生/突变型杂合子F/S(33/35.11%)、F/C(2/2.13%),突变型纯合子S/S(15/15.96%)、C/C(1/1.06%)和突变型杂合子S/C(4/4.26%)。各种群之间基因型比例各不相同。结论海口市kdr基因突变率较高,应进一步监测当地蚊虫抗药性,合理使用杀虫剂。
Objective To investigate the mutations in the knockdown resistance(kdr) gene of dengue vector Aedes albopictus in Haikou, China, as well as genotypes and their distribution in different areas, to analyze the influencing factors on insecticide resistance for scientific control of Ae. albopictus. Methods In June and July, 2018, the larvae and pupae of Ae.albopictus were collected from the urban and suburb areas of Haikou and reared to adults. The collected individuals were identified by morphology and were cryopreserved at -20 ℃ with anhydrous ethanol. DNA was extracted from a single mosquito, partial fragments of the voltage-gated sodium channel gene on nerve cell membrane were amplified by PCR, and sequencing was performed to analyze the mutations in the kdr gene. Results In the 94 Ae. albopictus mosquitoes in the east, south, west, north, and middle of Haikou, mutation was detected at the F1534 locus of the kdr gene. There were four alleles at the 1534 locus, namely, wild-type TTC/F(113/60.11%), mutant TCC/S(66/35.11%), mutant TCG/S(1/0.53%),and mutant TGC/C(8/4.26%); there were six genotypes, i.e., homozygous wild-type F/F(39/41.49%), wild/mutant heterozygote F/S(33/35.11%) and F/C(2/2.13%), homozygous mutant S/S(15/15.96%) and C/C(1/1.06%), and mutant heterozygote S/C(4/4.26%). The proportions of genotypes varied between populations. Conclusion The kdr mutation rate is high in Ae. albopictus in Haikou. Local mosquito resistance should be further monitored, so that insecticides are used appropriately.
Objective To investigate the mutations in the knockdown resistance(kdr) gene of dengue vector Aedes albopictus in Haikou, China, as well as genotypes and their distribution in different areas, to analyze the influencing factors on insecticide resistance for scientific control of Ae. albopictus. Methods In June and July, 2018, the larvae and pupae of Ae.albopictus were collected from the urban and suburb areas of Haikou and reared to adults. The collected individuals were identified by morphology and were cryopreserved at -20 ℃ with anhydrous ethanol. DNA was extracted from a single mosquito, partial fragments of the voltage-gated sodium channel gene on nerve cell membrane were amplified by PCR, and sequencing was performed to analyze the mutations in the kdr gene. Results In the 94 Ae. albopictus mosquitoes in the east, south, west, north, and middle of Haikou, mutation was detected at the F1534 locus of the kdr gene. There were four alleles at the 1534 locus, namely, wild-type TTC/F(113/60.11%), mutant TCC/S(66/35.11%), mutant TCG/S(1/0.53%),and mutant TGC/C(8/4.26%); there were six genotypes, i.e., homozygous wild-type F/F(39/41.49%), wild/mutant heterozygote F/S(33/35.11%) and F/C(2/2.13%), homozygous mutant S/S(15/15.96%) and C/C(1/1.06%), and mutant heterozygote S/C(4/4.26%). The proportions of genotypes varied between populations. Conclusion The kdr mutation rate is high in Ae. albopictus in Haikou. Local mosquito resistance should be further monitored, so that insecticides are used appropriately.
引文
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[8] Fernando SD,Hapugoda M,Perera R,et al. First report of V1016G and S989P knockdown resistant(kdr)mutations in pyrethroid-resistant Sri Lankan Aedes aegypti mosquitoes[J].Parasit Vectors,2018,11(1):526. DOI:10.1186/s13071-018-3113-0.
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[10]王晓花,陈辉莹,杨新艳,等.海口市白纹伊蚊对菊酯类杀虫剂的抗药性及击倒抗性基因突变分析[J].第二军医大学学报,2015,36(8):832-838. DOI:10.3724/SP.J.1008.2015.00832.
[11]国务院办公厅.国务院关于推进海南国际旅游岛建设发展的若干意见[国发[2009]44号][Z].(2010-01-04)[2018-11-16]. http://www.gov.cn/zwgk/2010-01/04/content_1502531.htm.
[12]吴维学,金玉明,孙莲英,等.海南省2006年登革热传播媒介监测分析[J].中国热带医学,2007,7(10):1863-1864,1920.DOI:10.3969/j.issn.1009-9727.2007.10.071.
[13]杜建伟,潘先海.中国登革热流行概况与流行特征[J].中华流行病学杂志,2010,31(12):1429-1433. DOI:10.3760/cma.j.issn.0254-6450.2010.12.025.
[14]罗永全,周艳华,王忠平,等.“双创”背景下海口市政府危机公关管理理念和能力建设的问题与对策研究[J].经济师,2017(1):173-176. DOI:10.3969/j.issn.1004-4914.2017.01.086.
[15] Kasai S,Ng LC,Lam-Phua SG,et al. First detection of a putative knockdown resistance gene in major mosquito vector,Aedes albopictus[J]. Jpn J Infect Dis,2011,64(3):217-221.
[16]高景鹏,李凯利,陈辉莹,等.菊酯类杀虫剂对海口市白纹伊蚊的效果测定[J].中国热带医学,2017,17(9):867-870.DOI:10.13604/j.cnki.46-1064/r.2017.09.04.
[17] Hasmiwati H,Rusjdi SR,Nofita E. Detection of Ace-1 gene with insecticides resistance in Aedes aegypti populations from DHFendemic areas in Padang,Indonesia[J]. Biodiversitas,2018,19(1):31-36. DOI:10.13057/biodiv/d190105.
[18] Feng XY,Yang C,Yang YC,et al. Distribution and frequency of G119S mutation in Ace-1 gene within Anopheles sinensis populations from Guangxi,China[J]. Malar J,2015,14(1):470. DOI:10.1186/s12936-015-1000-0.
[19] Ishak IH,Riveron JM,Ibrahim SS,et al. The Cytochrome P450gene CYP6P12 confers pyrethroid resistance in kdr-free Malaysian populations of the dengue vector Aedes albopictus[J].Sci Rep,2016,6(1):24707. DOI:10.1038/srep24707.
[2]杨舒然,刘起勇.白纹伊蚊的全球分布及扩散趋势[J].中国媒介生物学及控制杂志,2013,24(1):1-4.
[3]孟凤霞,王义冠,冯磊,等.我国登革热疫情防控与媒介伊蚊的综合治理[J].中国媒介生物学及控制杂志,2015,26(1):4-10. DOI:10.11853/j.issn.1003.4692.2015.01.002.
[4]陈翰明,高景鹏,姜进勇,等.我国白纹伊蚊现场群体击倒抗性基因I1532和F1534突变检测及I1532T突变等位基因报告[J].中国媒介生物学及控制杂志,2018,29(2):120-125.DOI:10.11853/j.issn.1003.8280.2018.02.002.
[5]姜进勇,陈辉莹,周红宁,等.云南省三带喙库蚊对DDT和溴氰菊酯抗性群体的击倒抗性基因突变分析[J].中国寄生虫学与寄生虫病杂志,2017,35(6):536-540.
[6]武松,马尔健,刘茜,等.中华按蚊击倒抗性突变PCR检测方法的建立[J].中国人兽共患病学报,2011,27(11):1008-1010. DOI:10.3969/j.issn.1002-2694.2011.11.014.
[7] Kristan M,Abeku TA,Lines J. Effect of environmental variables and kdr resistance genotype on survival probability and infection rates in Anopheles gambiae(s.s.)[J]. Parasit Vectors,2018,11(1):560. DOI:10.1186/s13071-018-3150-8.
[8] Fernando SD,Hapugoda M,Perera R,et al. First report of V1016G and S989P knockdown resistant(kdr)mutations in pyrethroid-resistant Sri Lankan Aedes aegypti mosquitoes[J].Parasit Vectors,2018,11(1):526. DOI:10.1186/s13071-018-3113-0.
[9] Gao JP,Chen HM,Shi H,et al. Correlation between adult pyrethroid resistance and knockdown resistance(kdr)mutations in Aedes albopictus(Diptera:Culicidae)field populations in China[J]. Infect Dis Poverty,2018,7(1):86. DOI:10.1186/s40249-018-0471-y.
[10]王晓花,陈辉莹,杨新艳,等.海口市白纹伊蚊对菊酯类杀虫剂的抗药性及击倒抗性基因突变分析[J].第二军医大学学报,2015,36(8):832-838. DOI:10.3724/SP.J.1008.2015.00832.
[11]国务院办公厅.国务院关于推进海南国际旅游岛建设发展的若干意见[国发[2009]44号][Z].(2010-01-04)[2018-11-16]. http://www.gov.cn/zwgk/2010-01/04/content_1502531.htm.
[12]吴维学,金玉明,孙莲英,等.海南省2006年登革热传播媒介监测分析[J].中国热带医学,2007,7(10):1863-1864,1920.DOI:10.3969/j.issn.1009-9727.2007.10.071.
[13]杜建伟,潘先海.中国登革热流行概况与流行特征[J].中华流行病学杂志,2010,31(12):1429-1433. DOI:10.3760/cma.j.issn.0254-6450.2010.12.025.
[14]罗永全,周艳华,王忠平,等.“双创”背景下海口市政府危机公关管理理念和能力建设的问题与对策研究[J].经济师,2017(1):173-176. DOI:10.3969/j.issn.1004-4914.2017.01.086.
[15] Kasai S,Ng LC,Lam-Phua SG,et al. First detection of a putative knockdown resistance gene in major mosquito vector,Aedes albopictus[J]. Jpn J Infect Dis,2011,64(3):217-221.
[16]高景鹏,李凯利,陈辉莹,等.菊酯类杀虫剂对海口市白纹伊蚊的效果测定[J].中国热带医学,2017,17(9):867-870.DOI:10.13604/j.cnki.46-1064/r.2017.09.04.
[17] Hasmiwati H,Rusjdi SR,Nofita E. Detection of Ace-1 gene with insecticides resistance in Aedes aegypti populations from DHFendemic areas in Padang,Indonesia[J]. Biodiversitas,2018,19(1):31-36. DOI:10.13057/biodiv/d190105.
[18] Feng XY,Yang C,Yang YC,et al. Distribution and frequency of G119S mutation in Ace-1 gene within Anopheles sinensis populations from Guangxi,China[J]. Malar J,2015,14(1):470. DOI:10.1186/s12936-015-1000-0.
[19] Ishak IH,Riveron JM,Ibrahim SS,et al. The Cytochrome P450gene CYP6P12 confers pyrethroid resistance in kdr-free Malaysian populations of the dengue vector Aedes albopictus[J].Sci Rep,2016,6(1):24707. DOI:10.1038/srep24707.