Objectives: Quantifying IgG antibodies to pertussis toxin (PT) is the most specific and sensitive method for the serodiagnosis of a Bordetella pertussis infection. Since PT is a component of acellular pertussis vaccines, anti-PT IgG is also induced by vaccination, precluding pertussis serodiagnosis based exclusively on anti-PT IgG in recently vaccinated subjects. Here, we aim to identify additional B. pertussis-specific serological markers that can discriminate between infection and recent vaccination. Methods: The clinical usefulness of measuring IgA directed to the vaccine antigen PT and IgG directed to non-vaccine antigens (Fim2/3, LPS, ACT, CatACT) was evaluated in nine well characterized subject groups, aged 10e89 years (n ¼ 390). Serum anti-PT IgG levels (>125 IU/mL) served as an indicator for a recent B. pertussis infection. Comparing symptomatic pertussis-infected subjects (n ¼ 140) with recently vaccinated, non-infected subjects (n ¼ 100) revealed the optimal cut-off, accuracy, sensitivity and specificity for each single parameter. Results: For pertussis diagnosis in recently vaccinated subjects, the measurement of anti-PT IgA (cut-off 15 IU/mL) and anti-ACT IgG (cut-off 15 U/mL) resulted in accuracies of 95% (91.5e97.1) and 87.5% (82.7 e91.1), sensitivities of 92.9% (87.4e96.0) and 83.6% (76.5e88.8) and specificities of 98% (93.0e99.4) and 93% (86.3e96.6), respectively. Comparing anti-PT IgA levels between the youngest (10e19 years, n ¼ 38) and oldest (70e89 years, n ¼ 17) age groups revealed an age-dependent increase in antibody levels in pertussis-infected subjects (p < 0.0001). Conclusions: Reflex testing of anti-PT IgA and anti-ACT IgG improves pertussis serodiagnosis in recently vaccinated symptomatic subjects with elevated anti-PT IgG levels. Furthermore, both markers can discriminate between vaccination and recent infection in pertussis serosurveillance studies.