Deteksi Virus Dengue Tipe 2 dengan Cara Hibridisasi in situ
(Detection of Tipe 2 Dengue Virus by in situ Hibridization)
MAKSUM RADJI, AMIN SOEBANDRIO, MIRAWATI SUDIRO & PRATIWI SUDARMONO
Bagian Mikrobiologi FK UI, Jln. Pegangsaan Timur No. 16, Jakarta 10320 Tel. 062-021-3100806, Faks. 062-021-3100810, Email: amin0207@rad.net.id
Demonstration of dengue virus (DV) in infected cell wouold enable elaboration of pathogenesis and pathophysiology of dengue Fever and dengue Haemorrhagic Fever. Molecular detection of DV would give high sensitivity as well as specifity. C6/36 mosquito cell line artificially infected with DV was used as a model of DV infected cell. 290-bp cDNA of envelope region of DV type 2 (DV-2) labeled with digoxigenin-11-dUTP was used as probe. Hybridization was performed directly to infected cell fixed on to glass slide (in situ). 10 ng/ul of the probe was able to detect as low as 10x TCID 50 infecting DV-2. The signal produced was not found in negative control and was clearly increasing in infecting viral dose dependent manner. There was no cross reactivity between DV-2 probe and DV-3 and vice versa. The DV-2 probe was sensitive yet specific in demonstrating the presence of DV-2 in infected cell.
Key word: hybridization, digoxygenin (DIG), pathogenesis
