Specific Primer Design for Acetylcholinesterase-1 (ace-1) as a Putative Insecticide Resistance Gene for Aedes aegypti

Abstract

The increase in cases of Dengue Hemorrhagic Fever (DHF) is attributed to an increase in DHF vectors, which is a
consequence of a failing mosquito control program. One major factor is specific gene resistance to insecticides. Research on resistance genes is crucial for effective vector control. One of the putative insecticide-resistant genes against Aedes aegypti mosquitoes is ace-1 (acetylcholinesterase-1), currently, there is no specific primer information that can amplify the ace-1 gene. This study aimed to design a specific primer for the ace-1 gene in Ae. aegypti. The study is an observational descriptive study in silico using NCBI, Primer3web, and BLAST databases, followed by an in vitro confirmation stage using the Polymerase Chain Reaction (PCR). The accession number of the ace-1 gene was MK896351.1. The research results showed two potential specific primer pairs; primer 1 with an amplicon length of ±413 bp and an annealing temperature of 49.6°C, while primer 2 with an amplicon length of ±417 bp and an annealing temperature of 51.3°C. The PCR product was visualized using 2% agarose gel and EtBr dye with gel documentation. In conclusion, this research successfully acquired two potential specific primer pairs for amplifying the ace-1 gene as a putative insecticide-resistant gene in Ae. aegypti mosquitoes.