Detection of Type 2 Dengue Virus by in situ Hybridization

Abstract

Demonstration of dengue virus DV in infected cell would enable elaboration of pathogenesis and pathoplsysiology of dengue Fever and dengue Haemorrhagic Fever. Molecular detection of DV would give high sensitivity as well as specifity. C6136 mosquito cell line artificially infected with DV was used as a model of DV infected cell. 290-bp eDNA of envelope region of DV type 2 DV-2 labeled with digoxigenin-il-dUT? 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 lOx TCJD 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.