Development and evaluation of an antibody capture ELISA for detection of IgG to Epstein-Barr virus in oral fluid samples

The development of an EBV IgG antibody capture ELISA (GACELISA) for detection of EBV viral capsid antigen specific IgG in oral fluids is described. The assay was optimised and evaluated using paired serum and oral fluid samples from healthy laboratory staff (n=82) and oral fluids collected either for routine measles, mumps, and rubella testing (n=629) or for an epidemiological study of atopic dermatitis (n=252). Statistical analysis by mixture modelling was used to determine the GACELISA cut-off and to estimate the sensitivity and specificity of the assay. Sensitivity and specificity was also assessed by comparing the results of immunofluorescence assay for EBV specific IgG in serum with those of GACELISA in 82 matching oral fluids. Compared to serum immunofluorescence assay, oral fluid GACELISA was found to have a sensitivity of 82.2 and 88.9% specificity with these samples. Mixture modelling, predicted the GACELISA to be 88.4% sensitive and of 99.4% specific. The prevalence of antibody to EBV in oral fluids was found to be 73.8% in laboratory staff, 34.4--73.9% in measles, mumps, and rubella patients and 22.2% in atopic dermatitis study participants. A robust, reliable and reproducible EBV GACELISA has been developed which will be a useful tool for epidemiological investigations.