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Journal of Clinical Microbiology, September 2005, p. 4602-4606, Vol. 43, No. 9
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.9.4602-4606.2005

Comparison of Methods for Detection of Vaccinia Virus in Patient Specimens

Daniel P. Fedorko,1* Jeanne C. Preuss,1 Gary A. Fahle,1 Li Li,1 Steven H. Fischer,1 Patricia Hohman,2 and Jeffrey I. Cohen2

Warren G. Magnuson Clinical Center,1 Laboratory of Clinical Infectious Diseases, National Institute for Allergy and Infectious Diseases, National Institutes of Health, U.S. Department of Health and Human Services, Bethesda, Maryland 208922

Received 4 March 2005/ Returned for modification 24 May 2005/ Accepted 20 June 2005

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.

* Corresponding author. Mailing address: Microbiology Service, DLM, Clinical Center, National Institutes of Health, Building 10, Room 2C385, 10 Center Drive, MSC 1508, Bethesda, MD 20892-1508. Phone: (301) 496-4433. Fax: (301) 402-1886. E-mail: dfedorko{at}nih.gov.

Journal of Clinical Microbiology, September 2005, p. 4602-4606, Vol. 43, No. 9
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.9.4602-4606.2005






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