Isolation of dengue viruses by intracerebral inoculation of mosquito larvae.

The isolation of dengue viruses from clinical specimens has always posed a particularly difficult problem. The use of invertebrate cell cultures such as AP-61 and C6/36 has reduced the time required for definitive diagnosis to within a week. More recently, inoculation of adult mosquitoes has been used but it requires more than a week to reach a confirmed laboratory diagnosis. We describe a method using intracerebral inoculation of immobilized fourth instar of Toxorhynchites splendens larvae for the isolation of dengue viruses from clinical specimens which yields results within a few days following incubation at 32 degrees C.