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.