Campylobacter jejuni is a causative pathogen of human acute bacterial gastroenteritis. Infected poultry products are regarded as a major source for human C. jejuni infection. The flagellar capping protein (FliD) is highly conserved among C. jejuni strains/isolates and is antigenic as analysed by immunoblot. In this study, we used the FliD protein as a probe to survey the prevalence of C. jejuni antibodies in chickens from two areas in the United States. A total of 394 samples were tested. Sera from layer breeders of 44–52 weeks of age tested 100% positive, while 4- to 6-week broilers from 22 premises showed 7–100% positivity. These results demonstrate that anti-FliD antibodies were prevalent in the poultry population in the areas of serum samples collected.