title: Comparison of methods for detection of vaccinia virus in patient specimens.
abstract: We analyzed a shell vial culture assay (SVA), real-time PCR, and a direct fluorescent antibody assay (DFA) for rapid detection of vaccinia virus from vaccination sites of Dryvax vaccine recipients. Of 47 samples assayed, 100% were positive by PCR, 89% were positive by SVA, and 40% were positive by DFA. DFA was limited by the need for adequate numbers of cells, with 32% of samples inadequate for interpretation. DFA performed better with specimens from patients who had not previously received the vaccine. PCR was positive for longer times postvaccination than was SVA. Infectious virus could be recovered after 45 min of acetone fixation of shell vial coverslips. Commercially available polyclonal antibodies cross-reacted with other orthopoxviruses and herpes simplex 1, but commercially available monoclonal antibodies were specific for vaccinia virus. In summary, PCR was the most sensitive test for detecting vaccinia virus in clinical specimens, while the DFA was the most rapid but the least sensitive test.
authors.affiliation: Warren G. Magnuson Clinical Center, National Institutes for Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-1508, USA. dfedorko@nih.gov
authors.forename: Daniel P
authors.initials: DP
authors.lastname: Fedorko
authors.forename: Jeanne C
authors.initials: JC
authors.lastname: Preuss
authors.forename: Gary A
authors.initials: GA
authors.lastname: Fahle
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.forename: Steven H
authors.initials: SH
authors.lastname: Fischer
authors.forename: Patricia
authors.initials: P
authors.lastname: Hohman
authors.forename: Jeffrey I
authors.initials: JI
authors.lastname: Cohen
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D019085:Fluorescent Antibody Technique, Direct; D006367:HeLa Cells; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012867:Skin / Q000166:cytology / Q000473:pathology / Q000821:virology*; D012899:Smallpox / Q000517:prevention & control; D012900:Smallpox Vaccine / Q000008:administration & dosage / Q000009:adverse effects*; D013048:Specimen Handling / Q000379:methods; D014611:Vaccination / Q000009:adverse effects; D014615:Vaccinia / Q000821:virology*; D014616:Vaccinia virus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D014776:Virus Cultivation
chemical_list: D000914:Antibodies, Viral; D012900:Smallpox Vaccine