Hubungan Frekuensi Gen Knock-Down Resistance (KDR) V1016G, V410L, dan F1534C dengan Tingkat Resistensi Populasi Aedes aegypti di Denpasar, Bali

 ABSTRACT Insecticides are used to reduce the number of Aedes aegypti mosquitoes, a vector of the dengue virus that causes dengue hemorrhagic fever in Indonesia, including in Bali. However, in recent years, the resistance of the Ae. aegypti mosquito to insecticides has been reported in Bali. In addition, Ae. aegypti is also identified to be resistant to several types of insecticides whose resistance mechanism is not yet clear. Several studies have stated that the KDR genes are one of the causes of resistance to pyrethroid insecticides. Therefore, a study was conducted to determine the frequency of the KDR genes in the Ae. aegypti population in Denpasar, Bali which had been previously shown resistance to permethrin (type of pyrethroid). Forty-three females Ae. aegypti specimens from bioassay testing using permethrin stored in tubes containing 100% ethanol were used in this study. Each mosquito is put in a different tube. The mosquito DNA was then extracted. Three KDR genes, V1016G, V410L, and F1534C, were analyzed using the quantitative-PCR melt curve method at the Biomolecular Laboratory, FKIK Unwar. In this study, the mutation frequency of 1016G was 98%, 410L was 50%, and 1534C was 3.4%. The 410L mutant gene was shared by all specimens, although only in one allele (heterozygous). Of the 43 specimens, 39 had homozygous mutant V1016G, heterozygous V410L, and wild typehomozygous F1534C. It indicated that the frequency of the 1016G and 410L mutant genes is quite high, while the 1534C gene is low. The combination of the KDR 1016G and 410L genes was the most dominant found in permethrin-resistant Ae. aegypti specimens. Therefore, it is assumed that these two KDR genes play a role in forming permethrin resistance in Ae. aegypti. ABSTRAK Insektisida digunakan untuk menurunkan jumlah nyamuk Aedes aegypti yang merupakan vektor virus dengue penyebab penyakit demam berdarah dengue di Indonesia, termasuk di Bali. Namun, dalam beberapa tahun terakhir, resistensi nyamuk Ae. aegypti terhadap insektisida dilaporkan sudah terjadi di Bali. Selain itu, Ae. aegypti juga dilaporkan resisten terhadap beberapa jenis insektisida yang mekanisme terjadinya resistensi belum diketahui. Beberapa penelitian menyebutkan bahwa gen KDR menjadi salah satu penyebab terjadinya mekanisme resistensi terhadap insektisida golongan peritroid. Oleh sebab itu, dilakukan penelitian untuk mengetahui frekuensi gen KDR pada populasi Ae. aegypti di Denpasar, Bali yang sebelumnya telah terbukti memiliki resistensi permetrin (insektisida golongan peritroid). Sebanyak 43 spesimen Ae. aegypti betina hasil dari pengujian bioassay yang tersimpan dalam tabung yang berisi etanol 100% digunakan dalam penelitian ini. Tiap nyamuk dimasukkan dalam tabung yang berbeda. Selanjutnya dilakukan ekstraksi DNA nyamuk. Tiga jenis gen KDR, yaitu V1016G, V410L, and F1534C, dianalisis dengan metode quantitative-PCR melt curve di Laboratorium Biomolekuler FKIK Unwar. Dalam penelitian ini ditemukan frekuensi mutasi 1016G sebesar 98%, 410L 50%, dan 1534C sebesar 3,4%. gen mutan 410L dimiliki oleh seluruh spesimen walaupun hanya pada salah satu alel (heterozigot). Dari 43 spesimen yang diteliti, terdapat 39 spesimen yang memiliki profil genotip V1016G homosigot mutan, V410L heterosigot, dan F1534C homosigot wild type. Sehingga dapat disimpulkan bahwa frekuensi gen mutan 1016G dan 410L cukup tinggi, sedangkan gen 1534C rendah. Kombinasi gen KDR  1016G dan 410L paling dominan ditemukan pada spesimen Ae. aegypti resisten permetrin. Kedua gen KDR diasumsikan berperan dalam pembentukan resistensi permetrin pada Ae. aegypti.