Estimation of Corynebacterium diphtheriae antitoxin in human sera: a comparison of an enzyme-linked immunosorbent assay with the toxin neutralisation test.

Serum samples from 101 individuals were titrated for Corynebacterium diphtheriae antitoxin by an IgG-specific enzyme-linked immunosorbent assay (ELISA) and a neutralisation test in tissue culture (TC). In some of the sera, the concentrations of antitoxin determined by the two assays were different; antitoxin values in these sera were titrated again by neutralisation tests in guinea pigs (GNT). Antitoxin concentrations of greater than 0.01 IU/ml by GNT partly corresponded to values obtained in both ELISA and TC. Only the values from TC agreed with lower GNT results. Heat inactivation of sera was investigated and rejected as a possible reason for the discrepancy in the results. ELISA can be used to detect levels of less than 0.1 IU/ml, although the accuracy below 0.01 IU/ml, often considered a protective level, is questionable. At higher levels ELISA was reproducible for the titration of diphtheria antitoxin in human sera and offers a useful alternative to both in-vivo assays and TC.