Detection of anti-arboviral immunoglobulin G by using a monoclonal antibody-based capture enzyme-linked immunosorbent assay

Monoclonal antibody (MAb)-based capture enzyme-linked immunosorbent assays (ELISAs) for the detection of anti-arboviral immunoglobulin G (IgG ELISAs) were developed for a comprehensive array of medically important arboviruses from the Alphavirus, Flavivirus, and Bunyavirus genera, Tests were optimiz ed and standardized so that maximum homology could be maintained among work ing parameters for the different viral agents, enabling a wide range of vir uses to be easily tested for at one time. MAbs were screened for suitabilit y as capture vehicles for antigens from the three genera, The final test co nfiguration utilized group-reactive MAbs eastern equine encephalitis virus 1A4B-6, dengue 2 virus 4G2, and La Crosse encephalitis virus 10G5.4 to capt ure the specific inactivated viral antigens, Serum IgG was detected by usin g alkaline phosphatase-conjugated anti-human IgG (Fc portion). A dilution o f 1:400 was chosen as the universal screening serum dilution, with endpoint titrations of serum samples testing positive eliminating occasional false- positive results. IgG ELISA results correlated with those of the standard p laque-reduction neutralization assays, As expected, some test cross-reactiv ity was encountered within the individual genera, and tests were interprete d within the context of these reactions. The tests were standardized for la boratory diagnosis of arboviral infections, with the intent that they be us ed in tandem with the corresponding IgM antibody-capture ELISAs.