Development of a recombinant enzyme-linked immunosorbent assay for detection of antibodies against Epstein-Barr virus nuclear antigens 2A and 2B.
The baculovirus expression system was used to produce full-length Epstein-Barr virus nuclear antigens (EBNAs) 2A and 2B. Recombinant baculoviruses that contained the EBNA-2A- and EBNA-2B-encoding sequences were constructed. The proteins were expressed in Spodoptera frugiperda SF-9 cells infected with the recombinant viruses and were characterized by using monoclonal and human polyclonal antibodies by immunoblotting and immunofluorescence techniques. Partially purified extracts of the EBNA-2A- and EBNA-2B-infected insect cells were used to establish a new enzyme-linked immunosorbent assay for the detection of antibodies against EBNA-2A and EBNA-2B. Preferential reactivity toward the type A or type B EBNA-2 protein was observed in 36% of serum specimens from Swiss patients with acute infectious mononucleosis and in 81% of Swiss patients with latent Epstein-Barr virus infection. Of the patients in the latter group, sera from 76% reacted preferentially with EBNA-2A, sera from 5% reacted preferentially with EBNA-2B, sera from 12% showed similar reactivities against both antigens, and sera from 7% were nonreactive.
Geertsen
R
R
Institute for Medical Virology, University of Zurich, Switzerland.
Espander-Jansson
A
A
Dobec
M
M
Price
P
P
Wunderli
W
W
Rymo
L
L
D000208:Acute Disease; D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D002353:Carrier State / Q000175:diagnosis; D004268:DNA-Binding Proteins / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D019309:Epstein-Barr Virus Nuclear Antigens; D006566:Herpesviridae Infections / Q000175:diagnosis* / Q000453:epidemiology; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D009036:Moths / Q000166:cytology; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011994:Recombinant Proteins / Q000276:immunology; D013557:Switzerland / Q000453:epidemiology; D017735:Virus Latency
D000914:Antibodies, Viral; D000956:Antigens, Viral; D004268:DNA-Binding Proteins; D019309:Epstein-Barr Virus Nuclear Antigens; D011994:Recombinant Proteins