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Pertussis is a highly contagious disease for which prompt, point-of-care diagnosis remains an unmet clinical need. Results from conventional test modalities (nucleic acid detection, serology, and culture) take hours to days. To overcome this challenge, we identified a new biomarker (tracheal colonization factor A, TcfA) for detection of  Bordetella pertussis  infection by lateral flow immunoassay (LFIA). We developed a library of 28 epitope-mapped monoclonal antibodies against TcfA and incorporated three antibodies into a LFIA. The LFIA did not cross-react with common bacterial or fungal organisms, but did react with nine distinct  B. pertussis  strains. The minimal linear epitope sequences targeted by the LFIA were conserved in 98% of 954  B. pertussis  isolates collected across 12 countries from 1949–2017. The LFIA’s limit of detection was 3.0 × 10 5  CFU/mL with  B. pertussis  cells in buffer, 6.2 × 10 5  CFU/mL with nasopharyngeal washes from a non-human primate model, and 2.3 ng/mL with recombinant TcfA. The LFIA reacted with patient nasopharyngeal swab specimens containing as few as 1.8 × 10 6   B. pertussis  genomes/mL and showed no false-positives. Rapid (< 20 min) LFIA detection of TcfA as a biomarker for  B. pertussis  infection is feasible and may facilitate early detection of pertussis.

Tracheal colonization factor A (TcfA) is a biomarker for rapid and specific detection of Bordetella pertussis