EARLY SERODIAGNOSIS OF ACUTE HUMAN CYTOMEGALOVIRUS-INFECTION BY ENZYME-LINKED-IMMUNOSORBENT-ASSAY USING RECOMBINANT ANTIGENS

DNA fragments from eight different reading frames of human cytomegalov irus (HCMV) were generated by PCR and subsequently cloned and expresse d in Escherichia coli in fusion with glutathione S-transferase. The re combinant viral antigens were evaluated in immunoblot analyses. The mo st reactive antigens were purified and further evaluated in ELISAs. Fo r this, sera from healthy blood donors and immunocompetent individuals with acute HCMV infection, and follow-up sera from transplant recipie nts with acute primary HCMV infection were used. The results of our ex periments indicate that only three particular recombinant polypeptides from two viral proteins are necessary for serodiagnosis. While a frag ment covering amino acids (aa) 495 to 691 of pp150 (150/1) was the mos t suitable antigen for the identification of infected individuals in g eneral, immunoglobulin M antibodies against the C-terminal parts of pp 150 (aa 862 to 1048; 150/7) and p52 (aa 297 to 433; 52/3) proved to be excellent serological markers to monitor acute HCMV infection. The se lected recombinant antigens enable the improvement of serodiagnosis of HCMV-related diseases, especially during the early stages of infectio n.