{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,19]],"date-time":"2026-02-19T23:36:49Z","timestamp":1771544209289,"version":"3.50.1"},"reference-count":35,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,9,26]],"date-time":"2021-09-26T00:00:00Z","timestamp":1632614400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,9,26]],"date-time":"2021-09-26T00:00:00Z","timestamp":1632614400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100013076","name":"national major science and technology projects of china","doi-asserted-by":"publisher","award":["2017ZX10303404"],"award-info":[{"award-number":["2017ZX10303404"]}],"id":[{"id":"10.13039\/501100013076","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100011229","name":"state key laboratory of integrated management of pest insects and rodents","doi-asserted-by":"crossref","award":["IPM1915"],"award-info":[{"award-number":["IPM1915"]}],"id":[{"id":"10.13039\/501100011229","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Parasites Vectors"],"published-print":{"date-parts":[[2021,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec>\n                <jats:title>Background<\/jats:title>\n                <jats:p><jats:italic>Anopheles sinensis<\/jats:italic> is a dominant vector for malaria transmission in Asian countries. Voltage-gated sodium channel (VGSC) mutation-mediated knock-down resistance (<jats:italic>kdr<\/jats:italic>) has developed in many <jats:italic>A. sinensis<\/jats:italic> populations because of intensive and long-term use of pyrethroids. Our previous study showed that multiple mutations at position 1014 of the VGSC were heterogeneously distributed in <jats:italic>A. sinensis<\/jats:italic> populations across Sichuan, China.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>To understand resistance genotypes at the haplotype level and reconstruct the phylogenetic relationship of <jats:italic>VGSC<\/jats:italic> haplotypes, a cost-effective next-generation sequencing (NGS)-based amplicon sequencing approach was established to clarify haplotypes containing codon 1014 of the <jats:italic>VGSC<\/jats:italic> gene from a total of 446 adults collected in 12 locations of Sichuan, China.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Nineteen (19) haplotypes were identified, including 11 wild 1014L, 6 resistance 1014F, and 2 resistance 1014C haplotypes. We found that resistance haplotypes of <jats:italic>A. sinensis VGSC<\/jats:italic> were widely distributed at frequencies ranging from 3.67 to 92.61%. The frequencies of the 1014C haplotype in the southeast of Sichuan (Luzhou, Guangan, and Suining) were relatively higher than those in other sampling locations. Phylogenetic analyses support that <jats:italic>kdr<\/jats:italic>-type mutation at position 1014 is not singly originated and resistance 1014C haplotypes evolve from TTT-encoding 1014F.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>A cost-effective next-generation sequencing (NGS)-based amplicon sequencing approach has been established in this study. The data revealed the patchy distribution of <jats:italic>VGSC<\/jats:italic> resistance haplotypes with overall high frequencies in Sichuan, China. Phylogenetic analyses support multiple origins and sequential evolution (1014L\u2009\u2192\u20091014F\u2009\u2192\u20091014C) for <jats:italic>kdr<\/jats:italic>-type mutations in <jats:italic>A. sinensis<\/jats:italic>.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Graphical abstract<\/jats:title>\n                \n              <\/jats:sec>","DOI":"10.1186\/s13071-021-05009-5","type":"journal-article","created":{"date-parts":[[2021,9,26]],"date-time":"2021-09-26T15:02:55Z","timestamp":1632668575000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Identification and phylogenetic analysis of voltage-gated sodium channel haplotypes in the malaria vector Anopheles sinensis using a high-throughput amplicon sequencing approach"],"prefix":"10.1186","volume":"14","author":[{"given":"Ruoyao","family":"Ni","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nian","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mei","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Weiping","family":"Qian","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2420-4705","authenticated-orcid":false,"given":"Xinghui","family":"Qiu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2021,9,26]]},"reference":[{"key":"5009_CR1","doi-asserted-by":"publisher","first-page":"569","DOI":"10.1146\/annurev-ento-112408-085423","volume":"55","author":"A Enayati","year":"2010","unstructured":"Enayati A, Hemingway J. Malaria management: past, present, and future. Annu Rev Entomol. 2010;55:569\u201391.","journal-title":"Annu Rev Entomol"},{"key":"5009_CR2","volume-title":"Integrated mosquito management","author":"B Lu","year":"1999","unstructured":"Lu B. Integrated mosquito management. 2nd ed. Beijing: Science Press; 1999.","edition":"2"},{"key":"5009_CR3","doi-asserted-by":"publisher","first-page":"807","DOI":"10.1016\/S0965-1748(97)00065-9","volume":"27","author":"TJ Smith","year":"1997","unstructured":"Smith TJ, Lee SH, Ingles PJ, Knipple DC, Soderlund DM. The L1014F point mutation in the house fly Vssc1 sodium channel confers knockdown resistance to pyrethroids. Insect Biochem Mol Biol. 1997;27:807\u201312.","journal-title":"Insect Biochem Mol Biol"},{"key":"5009_CR4","doi-asserted-by":"publisher","first-page":"513","DOI":"10.1124\/mol.104.006205","volume":"67","author":"J Tan","year":"2005","unstructured":"Tan J, Liu Z, Wang R, Huang ZY, Chen AC, Gurevitz M, et al. Identification of amino acid residues in the insect sodium channel critical for pyrethroid binding. Mol Pharmacol. 2005;67:513\u201322.","journal-title":"Mol Pharmacol"},{"key":"5009_CR5","doi-asserted-by":"publisher","first-page":"723","DOI":"10.1016\/j.ibmb.2011.05.004","volume":"41","author":"MJ Burton","year":"2011","unstructured":"Burton MJ, Mellor IR, Duce IR, Davies TG, Field LM, Williamson MS. Differential resistance of insect sodium channels with kdr mutations to deltamethrin, permethrin and DDT. Insect Biochem Mol Biol. 2011;41:723\u201332.","journal-title":"Insect Biochem Mol Biol"},{"key":"5009_CR6","doi-asserted-by":"publisher","first-page":"11785","DOI":"10.1073\/pnas.1305118110","volume":"110","author":"Y Du","year":"2013","unstructured":"Du Y, Nomura Y, Satar G, Hu Z, Nauen R, He SY, et al. Molecular evidence for dual pyrethroid-receptor sites on a mosquito sodium channel. Proc Natl Acad Sci U S A. 2013;110:11785\u201390.","journal-title":"Proc Natl Acad Sci U S A"},{"key":"5009_CR7","doi-asserted-by":"publisher","first-page":"6317","DOI":"10.1016\/j.febslet.2005.09.096","volume":"579","author":"PN Usherwood","year":"2005","unstructured":"Usherwood PN, Vais H, Khambay BP, Davies TG, Williamson MS. Sensitivity of the Drosophila para sodium channel to DDT is not lowered by the super-kdr mutation M918T on the IIS4-S5 linker that profoundly reduces sensitivity to permethrin and deltamethrin. FEBS Lett. 2005;579:6317\u201325.","journal-title":"FEBS Lett"},{"key":"5009_CR8","doi-asserted-by":"publisher","first-page":"305","DOI":"10.1085\/jgp.115.3.305","volume":"115","author":"H Vais","year":"2000","unstructured":"Vais H, Williamson MS, Goodson SJ, Devonshire AL, Warmke JW, Usherwood PN, et al. Activation of Drosophila sodium channels promotes modification by deltamethrin: reductions in affinity caused by knock-down resistance mutations. J Gen Physiol. 2000;115:305\u201318.","journal-title":"J Gen Physiol"},{"key":"5009_CR9","doi-asserted-by":"publisher","first-page":"243","DOI":"10.1146\/annurev-ento-011118-112420","volume":"64","author":"JG Scott","year":"2019","unstructured":"Scott JG. Life and death at the voltage-sensitive sodium channel: evolution in response to insecticide use. Annu Rev Entomol. 2019;64:243\u201357.","journal-title":"Annu Rev Entomol"},{"key":"5009_CR10","first-page":"783","volume":"30","author":"GJ Xu","year":"2014","unstructured":"Xu GJ, Li L, Yu T, Wu XH. Epidemic situation and progress in eliminating malaria in Sichuan. J Prev Med Inf. 2014;30:783\u20135.","journal-title":"J Prev Med Inf"},{"key":"5009_CR11","first-page":"75","volume":"35","author":"L Li","year":"2017","unstructured":"Li L, Liu Y, Xu GJ, Yu T, Zou Y, Wu XH, et al. Epidemiological characteristics of imported malaria in Sichuan Province in 2015. Chin J Parasitol Para Dis. 2017;35:75\u201385.","journal-title":"Chin J Parasitol Para Dis"},{"key":"5009_CR12","first-page":"321","volume":"73","author":"H Cheng","year":"1995","unstructured":"Cheng H, Yang W, Kang W, Liu C. Large-scale spraying of bednets to control mosquito vectors and malaria in Sichuan. China Bull World Health Organ. 1995;73:321\u20138.","journal-title":"China Bull World Health Organ"},{"key":"5009_CR13","doi-asserted-by":"publisher","first-page":"889","DOI":"10.1016\/j.pt.2017.06.010","volume":"33","author":"S Zhang","year":"2017","unstructured":"Zhang S, Guo S, Feng X, Afelt A, Frutos R, Zhou S, et al. Anopheles vectors in mainland China while approaching malaria elimination. Trends Parasitol. 2017;33:889\u2013900.","journal-title":"Trends Parasitol"},{"key":"5009_CR14","doi-asserted-by":"publisher","first-page":"169","DOI":"10.1186\/s13071-021-04662-0","volume":"14","author":"W Qian","year":"2021","unstructured":"Qian W, Liu N, Yang Y, Liu J, He J, Chen Z, et al. A survey of insecticide resistance-conferring mutations in multiple targets in Anopheles sinensis populations across Sichuan. China Parasit Vectors. 2021;14:169.","journal-title":"China Parasit Vectors"},{"key":"5009_CR15","doi-asserted-by":"publisher","first-page":"157","DOI":"10.1111\/j.1365-2583.2006.00620.x","volume":"15","author":"FD Rinkevich","year":"2006","unstructured":"Rinkevich FD, Zhang L, Hamm RL, Brady SG, Lazzaro BP, Scott JG. Frequencies of the pyrethroid resistance alleles of Vssc1 and CYP6D1 in house flies from the eastern United States. Insect Mol Biol. 2006;15:157\u201367.","journal-title":"Insect Mol Biol"},{"key":"5009_CR16","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0055475","volume":"8","author":"D Zhong","year":"2013","unstructured":"Zhong D, Chang X, Zhou G, He Z, Fu F, Yan Z, et al. Relationship between knockdown resistance, metabolic detoxification and organismal resistance to pyrethroids in Anopheles sinensis. PLoS ONE. 2013;8: e55475.","journal-title":"PLoS ONE"},{"key":"5009_CR17","doi-asserted-by":"publisher","first-page":"2114","DOI":"10.1093\/bioinformatics\/btu170","volume":"30","author":"AM Bolger","year":"2014","unstructured":"Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics. 2014;30:2114\u201320.","journal-title":"Bioinformatics"},{"key":"5009_CR18","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1186\/1471-2105-13-31","volume":"13","author":"AP Masella","year":"2012","unstructured":"Masella AP, Bartram AK, Truszkowski JM, Brown DG, Neufeld JD. PANDAseq: paired-end assembler for illumina sequences. BMC Bioinform. 2012;13:31.","journal-title":"BMC Bioinform"},{"key":"5009_CR19","doi-asserted-by":"publisher","first-page":"543","DOI":"10.1002\/ps.5543","volume":"76","author":"C D\u00e9lye","year":"2020","unstructured":"D\u00e9lye C, Michel S, Pernin F, Gautier V, Gislard M, Poncet C, et al. Harnessing the power of next-generation sequencing technologies to the purpose of high-throughput pesticide resistance diagnosis. Pest Manag Sci. 2020;76:543\u201352.","journal-title":"Pest Manag Sci"},{"key":"5009_CR20","doi-asserted-by":"publisher","first-page":"803","DOI":"10.1093\/jme\/tjz001","volume":"56","author":"S Chen","year":"2019","unstructured":"Chen S, Qin Q, Zhong D, Fang X, He H, Wang L, et al. Insecticide resistance status and mechanisms of Anopheles sinensis (Diptera: Culicidae) in Wenzhou, an important coastal port city in China. J Med Entomol. 2019;56:803\u201310.","journal-title":"J Med Entomol"},{"key":"5009_CR21","doi-asserted-by":"publisher","first-page":"92","DOI":"10.1186\/1756-3305-7-92","volume":"7","author":"Q Qin","year":"2014","unstructured":"Qin Q, Li Y, Zhong D, Zhou N, Chang X, Li C, et al. Insecticide resistance of Anopheles sinensis and An. vagus in Hainan Island, a malaria-endemic area of China. Parasit Vectors. 2014;7:92.","journal-title":"Parasit Vectors"},{"key":"5009_CR22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0002889","volume":"8","author":"X Chang","year":"2014","unstructured":"Chang X, Zhong D, Fang Q, Hartsel J, Zhou G, Shi L, et al. Multiple resistances and complex mechanisms of Anopheles sinensis mosquito: a major obstacle to mosquito-borne diseases control and elimination in China. PLoS Negl Trop Dis. 2014;8: e2889.","journal-title":"PLoS Negl Trop Dis"},{"key":"5009_CR23","doi-asserted-by":"publisher","first-page":"411","DOI":"10.1186\/s12936-016-1467-3","volume":"15","author":"C Yang","year":"2016","unstructured":"Yang C, Feng X, Huang Z, Li M, Qiu X. Diversity and frequency of kdr mutations within Anopheles sinensis populations from Guangxi. China Malar J. 2016;15:411.","journal-title":"China Malar J"},{"key":"5009_CR24","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1007\/BF01731581","volume":"16","author":"M Kimura","year":"1980","unstructured":"Kimura M. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol. 1980;16:111\u201320.","journal-title":"J Mol Evol"},{"key":"5009_CR25","doi-asserted-by":"publisher","first-page":"1547","DOI":"10.1093\/molbev\/msy096","volume":"35","author":"S Kumar","year":"2018","unstructured":"Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol. 2018;35:1547\u20139.","journal-title":"Mol Biol Evol"},{"key":"5009_CR26","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1093\/oxfordjournals.molbev.a026036","volume":"16","author":"HJ Bandelt","year":"1999","unstructured":"Bandelt HJ, Forster P, R\u00f6hl A. Median-joining networks for inferring intraspecific phylogenies. Mol Biol Evol. 1999;16:37\u201348.","journal-title":"Mol Biol Evol"},{"key":"5009_CR27","doi-asserted-by":"publisher","first-page":"5463","DOI":"10.1073\/pnas.74.12.5463","volume":"74","author":"F Sanger","year":"1977","unstructured":"Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA. 1977;74:5463\u20137.","journal-title":"Proc Natl Acad Sci USA"},{"key":"5009_CR28","doi-asserted-by":"publisher","first-page":"228","DOI":"10.1186\/s13071-016-1513-6","volume":"9","author":"X Chang","year":"2016","unstructured":"Chang X, Zhong D, Lo E, Fang Q, Bonizzoni M, Wang X, et al. Landscape genetic structure and evolutionary genetics of insecticide resistance gene mutations in Anopheles sinensis. Parasit Vectors. 2016;9:228.","journal-title":"Parasit Vectors"},{"key":"5009_CR29","doi-asserted-by":"publisher","first-page":"137","DOI":"10.1186\/s12936-015-0662-y","volume":"14","author":"HW Zhang","year":"2015","unstructured":"Zhang HW, Liu Y, Hu T, Zhou RM, Chen JS, Qian D, et al. Knockdown resistance of Anopheles sinensis in Henan province. China Malar J. 2015;14:137.","journal-title":"China Malar J"},{"key":"5009_CR30","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1186\/s12936-015-0644-0","volume":"14","author":"Y Wang","year":"2015","unstructured":"Wang Y, Yu W, Shi H, Yang Z, Xu J, Ma Y. Historical survey of the kdr mutations in the populations of Anopheles sinensis in China in 1996\u20132014. Malar J. 2015;14:120.","journal-title":"Malar J"},{"key":"5009_CR31","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0052761","volume":"7","author":"FD Rinkevich","year":"2012","unstructured":"Rinkevich FD, Hedtke SM, Leichter CA, Harris SA, Su C, Brady SG, et al. Multiple origins of kdr-type resistance in the house fly, Musca domestica. PLoS ONE. 2012;7: e52761.","journal-title":"PLoS ONE"},{"key":"5009_CR32","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.ibmb.2014.03.012","volume":"50","author":"K Dong","year":"2014","unstructured":"Dong K, Du Y, Rinkevich F, Nomura Y, Xu P, Wang L, et al. Molecular biology of insect sodium channels and pyrethroid resistance. Insect Biochem Mol Biol. 2014;50:1\u201317.","journal-title":"Insect Biochem Mol Biol"},{"key":"5009_CR33","doi-asserted-by":"publisher","first-page":"268","DOI":"10.1186\/1756-3305-6-268","volume":"6","author":"JC Lol","year":"2013","unstructured":"Lol JC, Castellanos ME, Liebman KA, Lenhart A, Pennington PM, Padilla NR. Molecular evidence for historical presence of knock-down resistance in Anopheles albimanus, a key malaria vector in Latin America. Parasit Vectors. 2013;6:268.","journal-title":"Parasit Vectors"},{"key":"5009_CR34","doi-asserted-by":"publisher","first-page":"450","DOI":"10.1186\/1756-3305-7-450","volume":"7","author":"AP Silva","year":"2014","unstructured":"Silva AP, Santos JM, Martins AJ. Mutations in the voltage-gated sodium channel gene of anophelines and their association with resistance to pyrethroids\u2014a review. Parasit Vectors. 2014;7:450.","journal-title":"Parasit Vectors"},{"key":"5009_CR35","doi-asserted-by":"publisher","first-page":"126","DOI":"10.1111\/imb.12206","volume":"25","author":"H Sun","year":"2016","unstructured":"Sun H, Tong KP, Kasai S, Scott JG. Overcoming super-knock down resistance (super-kdr) mediated resistance: multi-halogenated benzyl pyrethroids are more toxic to super-kdr than kdr house flies. Insect Mol Biol. 2016;25:126\u201337.","journal-title":"Insect Mol Biol"}],"container-title":["Parasites &amp; Vectors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13071-021-05009-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s13071-021-05009-5\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13071-021-05009-5.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,9,26]],"date-time":"2021-09-26T15:07:58Z","timestamp":1632668878000},"score":1,"resource":{"primary":{"URL":"https:\/\/parasitesandvectors.biomedcentral.com\/articles\/10.1186\/s13071-021-05009-5"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,9,26]]},"references-count":35,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2021,12]]}},"alternative-id":["5009"],"URL":"https:\/\/doi.org\/10.1186\/s13071-021-05009-5","relation":{},"ISSN":["1756-3305"],"issn-type":[{"value":"1756-3305","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,9,26]]},"assertion":[{"value":"18 June 2021","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 September 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"26 September 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Not applicable.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"We declare we have no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"499"}}