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Pseudomonas aeruginosa is associated with increased mortality in cystic fibrosis ( CF ) patients, and expresses type III secretion system proteins ( TTSP ), which is a common mechanism used by gram‐negative pathogens for delivery of anti‐host factors. Our aim was to investigate whether or not these antigens ( TTSP ) would be recognized by CF sera, by Western blot reaction. We have showed herein that all patients (n = 11) not chronically infected by P . aeruginosa had their first serum positive for TTSP ( ExoS , ExoT , PopB , and/or PopD ). All chronic patients had a strong positive serology to TTSP , although relatively weak reactions to TTSP were observed for some individuals in the negative control group. Therefore, TTSP that were early produced in P . aeruginosa infected CF patients, induced a detectable antibody response in those patients and were easily detected by Western‐blot reaction.