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Clinical and Vaccine Immunology, August 2008, p. 1259-1264, Vol. 15, No. 8
1071-412X/08/$08.00+0     doi:10.1128/CVI.00487-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Development of an Immunofluorescence Assay Using Recombinant Proteins Expressed in Insect Cells To Screen and Confirm Presence of Human Herpesvirus 8-Specific Antibodies{triangledown}

Veenu Minhas,1 Lynsey N. Crosby,1 Kay L. Crabtree,1 Saul Phiri,1 Tendai J. M'soka,2 Chipepo Kankasa,2 William J. Harrington,3 Charles D. Mitchell,3 and Charles Wood1*

Nebraska Center for Virology, School of Biological Sciences, University of Nebraska, Lincoln, Nebraska 68583,1 Department of Paediatrics and Child Health, University Teaching Hospital, Lusaka, Zambia,2 Department of Pediatrics, Miller School of Medicine, University of Miami, Miami, Florida, 331333

Received 11 December 2007/ Returned for modification 28 January 2008/ Accepted 20 May 2008

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

* Corresponding author. Mailing address: Nebraska Center for Virology, School of Biological Sciences, Room 102C, 4240 Fair Street, Morrison Center, University of Nebraska—Lincoln, Lincoln, NE 68583. Phone: (402) 472-4559. Fax: (402) 472-8722. E-mail: cwood1{at}unl.edu

{triangledown} Published ahead of print on 28 May 2008.

Clinical and Vaccine Immunology, August 2008, p. 1259-1264, Vol. 15, No. 8
1071-412X/08/$08.00+0     doi:10.1128/CVI.00487-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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