Sensitive detection of hantaviruses by biotin-streptavidin enhanced immunoassays based on bank vole monoclonal antibodies

Antigen capture enzyme-linked immunosorbent assays (ELISAs) based on detection of the viral nucleocapsid protein (N) were designed for rapid diagnosis of hantavirus infections. Several combinations of bank vole ( Clethrionomys glareolus) monoclonal antibodies with various N-epitope specificities were used for the development of two double-antibody sandwich forms of ELISA; PUU virus AG-ELISA for an exclusive detection of Puumala-related viruses, and Hantavirus AG-ELISA for a more extensive detection of all serotypes of hemorrhagic fever with renal syndrome (HFRS) viruses. The biotin-streptavidin system, in combination with horseradish peroxidase, rendered the assays' sensitivities similar to or even higher than immunoblotting. Calculated detection limits for the PUU virus and the Hantavirus AG-ELISAs were 405 and 50 focus forming units or 80 and 10 infected Vero E6 cells, respectively. The assays were evaluated and found to be suitable for convenient and routine detection of hantaviruses in cell cultures and in infected animal tissue.