Evaluation of optimized ELISA to detect anti-cytomegalovirus antibodies against immunogenic proteins

Herpesviruses are common viruses that have infected the vast majority of people around the globe. Accurate diagnosis of these viruses using cost-effective methods is important, especially for asymptomatic cases. Diagnostic differentiation between naturally occurring antibodies and other antibodies is an area that requires investigation. The aim of this study was to optimize an immunoassay to detect anti-herpesvirus antibodies directed against immunodominant proteins. A total of 120 sera were collected at two stages (62 samples for the pilot experiment during coronavirus pandemic, and 58 samples were collected recently). An ELISA test was developed using cytomegalovirus phosphoprotein 150 (pp150) and herpes simplex virus 2 glycoprotein D (HSV-2 gD) to test the specificity of the test. The cutoff value was 0.218, which indicates low background noise. There was competitive binding of the CMVpp150 as competitor compared to HSV-2 gD. The test can be reliably used to specifically determine anti-viral immunodominant proteins. Future studies should explore the binding of antibodies to other specific viral antigens to further expand the scale of testing. Furthermore, studies should be performed to reproduce these results with comparability using commercial kits.