Serum samples from 101 individuals were titrated for 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 >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 < 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.


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