HHV-8 ELISA based on a one-step affinity capture of biotinylated K8.1 antigen.
The immunogenic envelope antigen gp35-37 of human herpesvirus-8 (HHV-8) is encoded by orfK8.1. An ELISA is described using streptavidin capture of bacterially expressed and biotinylated recombinant K8.1 antigen. The antigen capture strategy provides a simple and reliable method, which does not require high yield production and purification of the recombinant antigen before the serological assay. The specificity and sensitivity of the K8.1 ELISA were validated by gp35-37 envelope antigen Western blot and anti-lytic membrane immunofluorescence assay using lytically induced HHV-8 infected BCBL-1 cells. Under the established ELISA conditions, eight of the 10 Kaposi's sarcoma patients and five of the 180 healthy blood donors had IgG antibodies to K8.1 envelope antigen.
Juhász
A
A
Department of Microbiology, Faculty of Medicine, University of Debrecen, Hungary, P.O. Box 17, 4032 Nagyerdei Blvd. 98, H-4012, Debrecen, Hungary. juhadr@jaguar.dote.hu
Kónya
J
J
Beck
Z
Z
Remenyik
E
E
Veress
G
G
Bégány
A
A
Medgyessy
I
I
Hunyadi
J
J
Gergely
L
L
D001710:Biotin; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D006023:Glycoproteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D019288:Herpesvirus 8, Human / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012514:Sarcoma, Kaposi / Q000097:blood / Q000821:virology*; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology
D006023:Glycoproteins; C113454:K8.1 protein, Human herpesvirus 8; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D001710:Biotin