Immunofluorescence (IF) and radioimmunoassay (RIA) have been compared as methods for detecting IgM antibody in 124 infants with confirmed or suspected congenital rubella. IF was used to test sucrose density gradient fractions and RIA to test fractions and whole serum.
When fractions were tested IF and RIA were equally specific and distinguished clearly between IgM and IgG, but RIA was the more sensitive method. The RIA titre in whole serum was always greater than in the peak IgM fraction and there was no evidence that testing the serum, rather than the fraction, could result in failure to detect IgM. With some sera RIA gave low titres which became negative after absorption with IgG-coated latex beads. The mechanism of this ‘false positive’ effect, which may have been due to IgM with anti-IgG activity, was not investigated, but if it can be removed by absorption it need not reduce the specificity of the test.
During the first 6 months of life IgM antibody was detected by RIA in 30 out of 32 unfractionated sera and by IF in fractions from 28 of these. After the age of 6 months IgM was found progressively less frequently and the greater sensitivity of RIA became a more obvious advantage: 17 out of 60 specimens were positive by RIA and 11 of these were negative by IF.
RIA testing of whole serum appears to be an economical, specific and sensitive method for detecting IgM antibody in congenital rubella, of particular value when the titre of antibody is low.