Infeksi Virus Dengue pada Nyamuk Aedes aegypti Menggunakan Artificial Blood Feeding dan Deteksi Virus Dengue Menggunakan Teknik Molekular

Abstract

Abstract. Artificial blood-feeding using the parafilm-M membrane can be used as an alternative solution and
substitute live animals as a source of blood. This method is not only be used for blood-feeding but also to infect
the dengue virus (DENV) to mosquitoes. This study was aimed to determine the effectiveness artificial blood
feeding using parafilm-M membrane in Aedes mosquitoes originated in Indonesia and determine the positivity
of mosquitoes infected by Indonesia DENV-1. DENV-1 was isolated from patient and propagated in Vero cell
culture. The feeding was done in cardboard cups after mosquitos have been starved for 4-17 hours before being
fed with human blood. A conical 50ml tube was prepared, and a hole was created in the tube lid. The tube
opening was covered with parafilm. Glycerol was added into conical tube and heated in water bath for an hour
at 55oC. A mixture of blood and DENV-1 was made with concentration of 10%. Detection of DENV in bloodfed
mosquitos was carried out by using Simplexa Dengue Real-Time RT-PCR assay. The results showed that
the prevalence of blood-fed mosquitoes reached 66.67% with fasting period for 17 hours. Blood feeding
mosquitoes are affected by duration of fasting period, blood-feeding time, and attractants from human skin
rubbed into parafilm-M membrane. The prevalence of blood-fed Ae. aegypti infected by DENV was 20.83%.
This study provides information on the effectiveness of artificial parafilm membrane blood-feeding in a
laboratory setting that will be useful for vector control study in Indonesia.