Utilizing Genomic Filter Paper for Direct Detection of V1016G Mutation on Aedes aegypti
Penulis/Author
dr. Taufik Mulya Perdana, M.Sc. (1); dr. Yogik Onky Silvana Wijaya, Ph.D. (2); dr. Alfin Harjuno Dwiputro, M.Sc. (3); Aesha Najla (4); Muhammad Rifqi Taftazani (5); Dini Aura Insani (6); Rachma Widya Pangesti (7); Dr.rer.nat. Risky Oktriani, S.Si., M.Biotech., M.Sc. (8); Prof. dr. Tri Baskoro Tunggul Satoto, M.Sc., Ph.D. (9)
Tanggal/Date
5 2024
Kata Kunci/Keyword
Abstrak/Abstract
Background: Several mutations on mosquitoes’ nerve voltage-gated sodium channel (VGSC) are linked to their resistance towards pyrethroid-based insecticide. The V1016G mutation is arguably the most important example of such mutation among Aedes aegypti population in Indonesia. Genomic surveillance of mosquito involved complicated technique, such as DNA isolation that may result in low DNA yields from sample. This can hinder the downstream processes including polymerase chain reaction (PCR). This study aimed to develop direct allele-specific PCR (AS-PCR) using an in-house filter paper as sample collection tools.
Methods: A total of 18 dried mosquito preparations and 2 isolated mosquito DNA samples were prepared for analysis. Specific primer for Ae. aegypti V1016G mutation was used. The size of produced amplicons will be 60 bp and 80 bp.
Results: Fifteen out of 18 dried mosquito preparations successfully generated clear amplicons at the 60 bp and 80 bp markers. Bands on the electrophoresis results indicated that AS-PCR results using in-house filter paper are comparable to the DNA isolation kit results. However, the amplicons from AS-PCR tend to have visible smudge.
Conclusion: AS-PCR from the in-house genomic filter paper was successfully used to detect V1016G mutation in Ae. aegypti, and it provides comparable results with conventional PCR using DNA templates isolated with DNA isolation kits. Further optimization is needed to produce clear bands on electrophoresis result.
