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

Authors

• Veenu Minhas, Nebraska Center for Virology, School of Biological Sciences, University of Nebraska-LincolnFollow
• Lynsey N. Crosby, Nebraska Center for Virology, School of Biological Sciences, University of Nebraska-Lincoln
• Kay L. Crabtree, Nebraska Center for Virology, School of Biological Sciences, University of Nebraska-Lincoln
• Saul Phiri, Nebraska Center for Virology, School of Biological Sciences, University of Nebraska-Lincoln
• Tendai J. M'soka, Department of Paediatrics and Child Health, University Teaching Hospital, Lusaka, Zambia
• Chipepo Kankasa, Department of Paediatrics and Child Health, University Teaching Hospital, Lusaka, Zambia
• William J. Harrington, Department of Pediatrics, Miller School of Medicine, University of Miami, Miami, Florida
• Charles D. Mitchell, Department of Pediatrics, Miller School of Medicine, University of Miami, Miami, FloridaFollow
• Charles Wood, Nebraska Center for Virology, School of Biological Sciences, University of Nebraska-LincolnFollow

Document Type

Article

Date of this Version

2008

Citation

CLINICAL AND VACCINE IMMUNOLOGY, Aug. 2008, p. 1259–1264,Vol. 15, No. 8

Comments

Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Abstract

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 antibodyenhanced 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.