Detection of Polymorphism on Voltage-gated Sodium Channel Gene of Indonesian Aedes aegypti Associated with Resistance to Pyrethroids

Budi Mulyaningsih, Sitti Rahmah Umniyati, Tri Baskoro Tunggul Satoto, Ernaningsih Ernaningsih, Dwi Aris Agung Nugrahaningsih


BACKGROUND: Aedes aegypti is a vector of several pathogens including dengue virus. Vector control is an effective way to break the transmission but unfortunately constant use of insecticides ultimately causes vector resistance. Pyrethroids have been used for about 15 years to combat Ae. aegypti in Yogyakarta Province, Indonesia. Single amino acid substitutions in the voltagegated sodium channel associated with pyrethroid resistance constitute one of the main causative factors of knockdown resistance (kdr). The aim of this study is to detect resistant status to cypermetrine and polymorphism on the voltagegated sodium channel gene of Ae. aegypti from 2 dengue endemic areas in Yogyakarta Province (Yogyakarta city and Sleman district).

METHODS: Pyrethroid resistance in Ae. aegypti mosquitoes was detected by using CDC Bottle Bioassay. To detect the polymorphism on the voltage-gated sodium channel gene of Ae. aegypti analyses were conducted by using PCR and direct DNA squencing with primers AaSCF1 and AaSCR4 for S989P, I1011M (or V), L1014F sites, and AaSCF7 and AaSCR7 for the F1534C site.

RESULTS: According to bioassay, the results for Ae. aegypti from Yogyakarta city (93% mortality) and Sleman district (88% mortality) suggest the possibility of resistance to cypermethrin. We observed polymorphism on voltagegated sodium channel gene on site F1534C (heterozygous).

CONCLUSION: The findings provide early evidence that the use of cypermethrin (pyrethroids) in Yogyakarta city and Sleman district, Yogyakarta Province, Indonesia is reducing its effectiveness to control Ae. aegypti. Recommendations include additional tests for confirmation.

KEYWORDS: Aedes aegypti, dengue virus, cypermethrin, Bioassay, Yogyakarta

Full Text:



World Health Organization. Dengue Guidelines for Diagnosis, Treatment, Prevention and Control: New Edition. Geneva: World Health Organization; 2009, article.

Scott TW, Morrison AC. Vector dynamics and transmission of dengue virus: implications for dengue surveillance and prevention strategies: vector dynamics and dengue preventn. Curr Top Microbiol Immunol. 2010; 338: 115-28, CrossRef.

Berge JB, Feyereisen R. Amichot M. Cytochrome P450 monooxygenases and insecticide resistance in insects. Philos Trans R Soc Lond Biol. 1998; 353: 1701-5, CrossRef.

Kawada H, Sai ZMO, Sein T, Emiko K, Yan NMM, Hlaing MT, et al. Co-occurrence of point mutations in the voltage-gated sodium channel of pyrethroid-resistant Aedes aegypti populations in Myanmar. PLoS Negl Trop Dis. 2014; 8: e3032, CrossRef.

Kawada H. An inconvenient truth of pyrethroid – does it have a promising future? In: Clark J, Bloomquist JR, Kawada H, editors. Advances in Human Vector Control (ACS Symposium Book 1014). New York: American Chemical Society; 2009. p.171-90, CrossRef.

Brengues C, Hawkes NJ, Chandre F, McCaroll L, Duchon S, et al. Pyrethroid and DDT cross-resistance in Aedes aegypti is correlated with novel mutations in the voltage-gated sodium channel gene. Med Vet Entomol, 2003; 17: 87-94, CrossRef.

Saavedra-Rodriguez K, Urdaneta-Marquez L, Rajatileka S, Moulton M, Flores AE, et al. A mutation in the voltage-gated sodium channel gene associated with pyrethroid resistance in Latin American Aedes aegypti. Insect Mol Biol, 2007; 16: 785-98, CrossRef.

Chang C, Shen WK, Wang TT, Lin YH, Hsu EL, et al. A novel amino acid substitution in a voltage-gated sodium channel is associated with knockdown resistance to permethrin in Aedes aegypti. Insect Biochem Mol Biol. 2009; 39: 272-8, CrossRef.

Ghiffari A, Fatimi H, Anwar C. Detection of insecticide synthetic pyrethroid resistance on dengue vector Aedes aegypti (L.) in Palembang using Polymerase Chain Reaction. Aspirator. 2013; 5: 37-44, article.

Yanola J, Somboon P, Prapanthadara L. A novel point mutation in the Aedes aegypti voltage-gated sodium channel gene associated with permethrin resistance. In: The 2nd International Conference on Dengue and Dengue Haemorhagic Fever, Oct. 15–17, 2008. Phuket, Thailand.

Yanola J, Somboon P, Walton C, Nachaiwieng W, Somwang P, et al. High-throughput assays for detection of the F1534C mutation in the voltagegated sodium channel gene in permethrin-resistant Aedes aegypti and the distribution of this mutation throughout Thailand. Trop Med Int Health, 2011; 16: 501-9, CrossRef.

Harris AF, Rajatileka S, Ranson H. Pyrethroid resistance in Aedes aegypti from Grand Cayman. Am J Trop Med Hyg 2010; 83: 277-84, CrossRef.

Srisawat R, Komalamisra N, Eshita Y, Zheng M, Ono K, et al. Point mutations in domain II of the voltage-gated sodium channel gene in deltamethrin-resistant Aedes aegypti (Diptera: Culicidae). Appl Entomol Zool, 2010; 45: 275–282, CrossRef.

Center for Diseases Control and Prevention. Guideline for Evaluating Insecticide Resistance in Vectors using the CDC Bottle Bioassay. Atlanta: Center for Diseases Control and Prevention; 2010, article.

Cui F, Raymond M, Qiao CL. Insecticide resistance in vector mosquitoes in China. Pest Manag Sci. 2006; 62: 1013-22, CrossRef.

Hemingway J, Hawkes NJ, McCarroll L, Ranson L. The molecular basis of insecticide resistance in mosquitoes. Insect Biochemistry and Molecular Biology. 2004; 34: 653-65, CrossRef.

Srisawat R, Komalamisra N, Eshita Y, Zheng M, Ono K, et al. Point mutations in domain II of the voltage-gated sodium channel gene in deltamethrin-resistant Aedes aegypti (Diptera: Culicidae). Appl Entomol Zool. 2010; 45: 275-82, CrossRef.

Aung TT, Win S, Aung S. Status report on epidemiology of dengue/dengue haemorrhagic fever in Myanmar. Dengue Bull. 1995; 20: 41-5.

Nauen R. Insecticide resistance in disease vectors of public health importance. Pest Manag Sci. 2007; 63: 628-33, CrossRef.

Kawada H. New mosquito control techniques as countermeasures against insecticide resistance. In: Perveen F, editor. Insecticides - Advances in Integrated Pest Management. London: InTech; 2012. p.657-82, CrossRef.


Indexed by:






The Prodia Education and Research Institute