The use of Toxorhynchites splendens for identification and quantitation ofserotypes contained in the tetravalent live attenuated dengue vaccine

Assurance of identity and quantity is an indispensable part of quality cont rol in the manufacture of vaccines. Dengue-1 PDK13, dengue-2 PDK53, dengue- 3 PGMK30F3 and dengue-4 PDK48 in the live attenuated tetravalent dengue vac cine were assayed by identification and quantitation in a mosquito system ( Toxorhynchites splendens). Each serotype of dengue virus was identified by dengue specific monoclonal antibodies in the indirect fluorescent antibody test. Virus content was estimated by calculating the 50% mosquito infectiou s dose (MID50). Differences from 0 to +/-0.5 log(10) were observed between the original monovalent titer and that from the blend which showed no signi ficant difference at 95% confidence limit (P < 0.05). This result indicates that there is no interference between dengue serotypes in mosquitoes infec ted by intrathoracic inoculation with the virus mixture. It can be also con cluded that this mosquito system can be used as an effective measure for in fectivity titration of each component in the tetravalent dengue vaccine. (C ) 1999 Elsevier Science Ltd. All rights reserved.