Development of an immunofluorescence assay using recombinant proteins expressed in insect cells to screen and confirm presence of human herpesvirus 8-specific antibodies.
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.
Nebraska Center for Virology, School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583, USA.
Veenu
V
Minhas
Lynsey N
LN
Crosby
Kay L
KL
Crabtree
Saul
S
Phiri
Tendai J
TJ
M'soka
Chipepo
C
Kankasa
William J
WJ
Harrington
Charles D
CD
Mitchell
Charles
C
Wood
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000276:immunology / Q000821:virology; D019288:Herpesvirus 8, Human / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D012680:Sensitivity and Specificity; D018411:Spodoptera; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D000914:Antibodies, Viral; D000956:Antigens, Viral; D006023:Glycoproteins; D007074:Immunoglobulin G; C113454:K8.1 protein, Human herpesvirus 8; C119019:ORF65 protein, human herpesvirus 8; D011994:Recombinant Proteins; D014764:Viral Proteins