Evaluation of Commercial Immunoassays for Rubella Virus IgG Detection in Low-Antibody Sera Using a Recombinant...

Rubella virus (RV) IgG quantification is essential for verifying immunity, particularly in prenatal care. However, substantial variability exists among commercial immunoassays, especially when testing low-antibody sera. In this study, we evaluated five commercial assays-four chemiluminescent immunoassays (CLIAs) and one Enzyme-linked Immunosorbent Assay (ELISA)-using a recombinant immunoblot (IB) as the reference method. A panel of 137 serum samples with low or undetectable IgG levels was analyzed. Sensitivity ranged from 19.6% to 70.1%, while specificity exceeded 94%. Only 18.6% of immunoblot-positive samples tested positive across all assays. Marked quantitative differences were observed, with the Atellica assay yielding the highest titers and Alinity the lowest. Reclassifying equivocal results as positive improved concordance without compromising specificity. These findings suggest that current cut-off values, derived from post-infection sera, may be inadequate for vaccinated populations. A single universal threshold may lead to misclassification and underestimation of immunity. Harmonization of assay calibrations, antigenic targets, and interpretation criteria is urgently needed to ensure reliable rubella immunity assessments in clinical and public health settings.