A NOVEL MU-CAPTURE ENZYME-LINKED-IMMUNOSORBENT-ASSAY BASED ON RECOMBINANT PROTEINS FOR SENSITIVE AND SPECIFIC DIAGNOSIS OF HEMORRHAGIC-FEVER WITH RENAL SYNDROME

Hantavirus nucleocapsid protein has recently been identified as a majo r antigen inducing an early and long-lasting humoral immune response i n patients with hemorrhagic fever with renal syndrome. A mu-capture en zyme-linked immunosorbent assay utilizing recombinant nucleocapsid pro teins of Hantavirus strains Hantaan 76-118 (Hantaan serotype) and CG 1 8-20 (Puumala serotype) as diagnostic antigens and specific monoclonal antibodies as the detection system has been developed. Histidine-tail ed recombinant proteins were expressed in Escherichia coli and purifie d in a single step by affinity chromatography on a nickel-chelate resi n. The assay was evaluated with a panel of sera from patients with hem orrhagic fever with renal syndrome originating from various geographic regions. The overall sensitivity of the mu-capture enzyme-linked immu nosorbent assay (both recombinant antigens) was 100%, and its specific ity was also found to be 100%. Immunoglobulin M antibodies were detect ed as early as on day 3, and maximum titers were obtained between days 8 and 25 after onset of the disease. The assay was regularly found to be positive within 3 to 4 months but in some cases up to 2 years afte r the acute phase of the disease.