Detection and identification of Variola virus in fixed human tissue after prolonged archival storage.
Smallpox disease has been eradicated from the human population since 1979, but is again a concern because of its potential use as an agent of bioterrorism or biowarfare. World Health Organization-sanctioned repositories of infectious Variola virus are known to occur in both Russia and the United States, but many believe other undeclared and unregulated sources of the virus could exist. Thus, validation of improved methods for definitive identification of smallpox virus in diagnostic specimens is urgently needed. In this paper, we describe the discovery of suspected Variola infected human tissue, fixed and preserved for decades in largely unknown solutions, and the use of routine histology, electron microscopy, and ultimately DNA extraction and fluorogenic 5' nuclease (TaqMan) assays for its identification and confirmation.
Schoepp
Randal J
RJ
Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Frederick, MD 21702-5011, USA. randal.schoepp@amedd.army.mil
Morin
Michelle D
MD
Martinez
Mark J
MJ
Kulesh
David A
DA
Hensley
Lisa
L
Geisbert
Thomas W
TW
Brady
Daniel R
DR
Jahrling
Peter B
PB
D001109:Archives; D001431:Bacteriological Techniques; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D005456:Fluorescent Dyes; D006801:Humans; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012867:Skin / Q000473:pathology / Q000821:virology; D012899:Smallpox / Q000175:diagnosis* / Q000821:virology; D019914:Taq Polymerase / Q000235:genetics / Q000378:metabolism; D016707:Tissue Fixation; D012901:Variola virus / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure
D004279:DNA, Viral; D005456:Fluorescent Dyes; D019914:Taq Polymerase