A sample of 643 healthy subjects from central Italy aged 20 to 80, were screened for diphtheria antitoxin. Serum diphtheria antitoxin was assayed by a new passive haemagglutination technique using turkey red blood cells sensitized with diphtheria toxoid, after having performed a correlation study between this technique and the reference in vivo neutralization test. Of the studied population 26.7% showed a lack of serum antitoxin titres considered to be protective. The rate of susceptible subjects increased with age, showing the highest value (38.9%) in the sixth decade of age. Males proved less protected than females; 53.2% of the male population aged 50–59 were lacking a protective anti-diphtheria immunity. On the basis of present results, a periodical revaccination of the entire adult population with reduced doses of diphtheria toxoid would be advisable.