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Clin. Vaccine Immunol. doi:10.1128/CVI.00487-07
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Development of an immunofluorescent assay using recombinant proteins expressed in insect cells for the screening and confirmation of human herpesvirus 8 antibodies

Nebraska Center for Virology, School of Biological Sciences, University of Nebraska Lincoln, Lincoln, Nebraska 68583 USA; Department of Paediatrics and Child Health, University Teaching Hospital, Lusaka, Zambia; Department of Pediatrics, Miller School of Medicine, University of Miami, Miami, Florida, 33133 USA

^* To whom correspondence should be addressed. Email: cwood1{at}unl.edu.

	Abstract

Human herpesvirus-8 (HHV-8) or Kaposi's sarcoma-associated herpesvirus (KSHV) has been linked to all forms of Kaposi's sarcoma (KS). Most current serological assays to detect HHV-8 antibodies have low concordance amongst themselves. To establish a sensitive and specific testing strategy to screen for HHV-8 antibodies, three HHV-8 proteins, ORF65, ORF73 and K8.1A, were expressed using baculoviral vectors in insect cells and incorporated into an monoclonal-enhanced immunofluorescence assay (mIFA) termed Sf9 3-antigen mIFA. The results with this mIFA were compared to those obtained with a standard mIFA utilizing a HHV-8 infected B-cell line (BC3 mIFA). Test sera were obtained from patients with diagnosed KS, HIV-1 infected patients at high risk for HHV-8 infection, and healthy controls from a local blood bank. Combined use of both assays together had a sensitivity of 94% and a specificity of 96%. The performance of these two assays when used together indicates that they may be useful for reliable detection of HHV-8 IgG antibodies in a population.