Specificity and sensitivity of the IgM capture immunoassay: studies of possible factors inducing false positive or false negative results.

The specificity and sensitivity of the IgM-capture immunoassay (IgM-CI) were evaluated for detection of rubella specific IgM and hepatitis B core (HBc) specific IgM. For rubella specific IgM, antibodies bound to the solid phase were detected by haemadsorption and for HBc specific IgM, by using HBc antigen (HBcAg) and radiolabelled IgG anti-HBc. Rheumatoid factor (RF) was found to interfere in the test for HBc specific IgM because IgM-RF bound to the solid phase reacted with aggregated radiolabelled HBc specific IgG. This false positive reaction did not occur when radiolabelled F(ab')2 was used instead of the whole IgG molecule. HBcAg purified from biological fluids might be coated with host IgG and under these conditions, HBcAg could react with RF. It was also demonstrated that high levels of IgG antibodies could interfere with IgG anti-mu coated-surface by means of non-immunological protein-protein interactions. In fact, IgG did not interfere in the rubella assay, whereas it did in the very sensitive anti-HBc test. To prevent this false-positive reaction, different dilution media were tested. Only the addition of non-specific IgG and fetal calf serum (FCS), to the dilution medium, seems to improve the specificity of the test. Furthermore, in order to decrease this non-specific IgG-IgG interaction and an occasional prozoning phenomenon, the dilution of serum to be tested was taken into account. Parameters considered to decrease sensitivity were also studied. RF, anti-F(ab')2 antibodies and non-specific IgM did not decrease significantly the sensitivity of the assay.