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 Epst ein-Barr virus nuclear antigens (EBNAs) 2A and 2B. Recombinant baculov iruses 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 b y using monoclonal and human polyclonal antibodies by immunoblotting a nd immunofluorescence techniques. Partially purified extracts of the E BNA-2A- and EBNA-2B-infected insect cells were used to establish a new enzyme-linked immunosorbent assay for the detection of antibodies aga inst 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 Swi ss patients with acute infectious mononucleosis and in 81% of Swiss pa tients with latent Epstein-Barr virus infection. Of the patients in th e latter group, sera from 76% reacted preferentially with EBNA-2A, ser a from 5% reacted preferentially with EBNA-2B, sera from 12% showed si milar reactivities against both antigens, and sem from 7% were nonreac tive.