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

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.