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      First identification of kdr allele F1534S in VGSC gene and its association with resistance to pyrethroid insecticides in Aedes albopictus populations from Haikou City, Hainan Island, China

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          Aedes albopictus is distributed widely in China, as a primary vector of Dengue fever and Chikungunya fever in south of China. Chemical insecticide control is one of the integrated programmes to prevent mosquito-borne diseases. Long-term applications of pyrethroids have resulted in the development of resistance in Ae. albopictus populations in China. However, the susceptibility of Ae. albopictus to pyrethroids in Hainan Island was unclear. Knockdown resistance (kdr), caused by point mutations in the VGSC gene, is one of the mechanisms that confer resistance to DDT and pyrethroids. This study was to investigate the resistance level of Ae. albopictus populations in Haikou City to three pyrethroid insecticides, and elucidate the relationship between the resistant phenotype and kdr mutations.


          The Aedes albopictus samples were collected in Xinbu Island (XI), Longtang Town (LT), Shishan Town (ST), Baishamen Park (BP), and Flower Market (FM) from Haikou City, Hainan Island, China. The larval susceptibility to deltamethrin, permethrin and beta-cypermethrin was tested by larval bioassays, and adult susceptibility to deltamethrin and DDT was determined by adult bioassays. The degree of resistance was determined by resistance ratio value (RR 50 > 3) for larvae and by mortality for adult. The kdr alleles at codon 1534 of the VGSC gene were genotyped. The relationship between kdr genotypes and resistant phenotypes was analyzed by Chi-square test.


          Out of five populations, assessed by larval bioassays, XI was susceptible to deltamethrin and permethrin; LT was susceptible to permethrin and beta-cypermethrin; and ST was susceptible to permithrin. FM and BP both were resistant to all of the three pyrethroids, and FM showed the highest degree of resistance, with RR 50 values from 65.17 to 436.36. A total of 493 individuals from the larval bioassays were genotyped for kdr alleles. Five alleles were detected, including two wildtype alleles, TTC(F) (67.04 %) and TTT(F) (0.41 %), and three mutant alleles, TGC(C) (0.30 %), TCC(S) (31.54 %) and TTG(L) (0.71 %). There was a clear correlation between mutant alleles (or F1534S) and resistant phenotypes ( P < 0.01).


          Two novel kdr mutant alleles F1534S and F1534L were detected in the pyrethroid resistant populations of Ae. albopictus in Haikou Hainan, China. For the first time, the mutant F1534S was associated with pyrethroid resistance in Ae. albopictus.

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          The online version of this article (doi:10.1186/s40249-016-0125-x) contains supplementary material, which is available to authorized users.

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          Most cited references 54

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          Insecticide resistance in insect vectors of human disease.

          Insecticide resistance is an increasing problem in many insect vectors of disease. Our knowledge of the basic mechanisms underlying resistance to commonly used insecticides is well established. Molecular techniques have recently allowed us to start and dissect most of these mechanisms at the DNA level. The next major challenge will be to use this molecular understanding of resistance to develop novel strategies with which we can truly manage resistance. State-of-the-art information on resistance in insect vectors of disease is reviewed in this context.
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            Molecular characterization of pyrethroid knockdown resistance (kdr) in the major malaria vector Anopheles gambiae s.s.

            Pyrethroid-impregnated bednets are playing an increasing role for combating malaria, especially in stable malaria areas. More than 90% of the current annual malaria incidence (c. 500 million clinical cases with up to 2 million deaths) is in Africa where the major vector is Anopheles gambiae s.s. As pyrethroid resistance has been reported in this mosquito, reliable and simple techniques are urgently needed to characterize and monitor this resistance in the field. In insects, an important mechanism of pyrethroid resistance is due to a modification of the voltage-gated sodium channel protein recently shown to be associated with mutations of the para-type sodium channel gene. We demonstrate here that one of these mutations is present in certain strains of pyrethroid resistant A. gambiae s.s. and describe a PCR-based diagnostic test allowing its detection in the genome of single mosquitoes. Using this test, we found this mutation in six out of seven field samples from West Africa, its frequency being closely correlated with survival to pyrethroid exposure. This diagnostic test should bring major improvement for field monitoring of pyrethroid resistance, within the framework of malaria control programmes.
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              A mutation in the voltage-gated sodium channel gene associated with pyrethroid resistance in Latin American Aedes aegypti.

              Pyrethroids are commonly used as mosquito adulticides and evolution of resistance to these compounds is a major threat to public health. 'Knockdown resistance' to pyrethroids (kdr) is frequently caused by nonsynonymous mutations in the voltage-gated sodium channel transmembrane protein (para) that reduce pyrethroid binding. Early detection of kdr is critical to the development of resistance management strategies in mosquitoes including Aedes aegypti, the most prevalent vector of dengue and yellow fever viruses. Brengues et al. described seven novel mutations in hydrophobic segment 6 of domain II of para in Ae. aegypti. Assays on larvae from strains bearing these mutations indicated reduced nerve sensitivity to permethrin inhibition. Two of these occurred in codons Iso1011 and Val1016 in exons 20 and 21 respectively. A transition in the third position of Iso1011 encoded a Met1011 replacement and a transversion in the second position of Val1016 encoded a Gly1016 replacement. We have screened this same region in 1318 mosquitoes in 32 additional strains; 30 from throughout Latin America. While the Gly1016 allele was never detected in Latin America, we found two new mutations in these same codons. A transition in the first position of codon 1011 encodes a Val replacement while a transition in the first position of codon 1016 encodes an Iso replacement. We developed PCR assays for these four mutations that can be read either on an agarose gel or as a melting curve. Selection experiments, one with deltamethrin on a field strain from Santiago de Cuba and another with permethrin on a strain from Isla Mujeres, Mexico rapidly increased the frequency of the Iso1016 allele. Bioassays of F(3) offspring arising from permethrin susceptible Val1016 homozygous parents and permethrin resistant Iso1016 homozygous parents show that Iso1016 segregates as a recessive allele in conferring kdr. Analysis of segregation between alleles at the 1011 and 1016 codons in the F(3) showed a high rate of recombination even though the two codons are only separated by a ~250 bp intron. The tools and information presented provide a means for early detection and characterization of kdr that is critical to the development of strategies for resistance management.

                Author and article information

                Infect Dis Poverty
                Infect Dis Poverty
                Infectious Diseases of Poverty
                BioMed Central (London )
                2 May 2016
                2 May 2016
                : 5
                [ ]Department of Tropical Infectious Diseases, Faculty of Tropical Medicine and Public Health, Second Military Medical University, Shanghai, 200433 China
                [ ]Haikou Center for Disease Control and Prevention, Haikou, 571100 China
                [ ]CDC Key Laboratory of Surveillance and Early-Warning on Infectious Disease, Haikou, 571100 China
                © Chen et al. 2016

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( applies to the data made available in this article, unless otherwise stated.

                Funded by: National Natural Sciences Foundation of China
                Award ID: 81371848
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                Research Article
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                © The Author(s) 2016

                china, aedes albopictus, pyrethroids, resistance, kdr mutation


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