Use of enzyme immunoassay and nucleic acid hybridization for detecting Sindbis virus in infected mosquitoes.
Aedes aegypti mosquitoes were inoculated intrathoracically with prototype Sindbis virus, held at 26.7 degrees C for from 0-95 h and placed at -70 degrees C. Individual mosquitoes were tested for virus by plaque assay in Vero cells, for viral RNA by nucleic acid hybridization using a cloned cDNA probe, and for viral protein by enzyme-linked immunosorbent assay. Virus was detected by plaque assay as early as 8 h after infection. Sindbis virus RNA was detected by nucleic acid hybridization 18 h after infection and by enzyme-linked immunosorbent assay 10 h after infection. The results of these comparisons suggest that both nucleic acid hybridization and enzyme-linked immunosorbent assay are applicable to direct detection of Sindbis virus in mosquitoes containing virus at levels usually found during arbovirus epidemics.
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis*; D009033:Culicidae / Q000382:microbiology*; D004797:Enzyme-Linked Immunosorbent Assay; D008297:Male; D009693:Nucleic Acid Hybridization; D012367:RNA, Viral / Q000032:analysis*; D012845:Sindbis Virus / Q000276:immunology / Q000302:isolation & purification*
D000956:Antigens, Viral; D012367:RNA, Viral