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Understanding the colonization of <i>Pseudomonas aeruginosa</i> (<i>P. aeruginosa</i>) in healthy humans is useful for future prevention and treatment of <i>P. aeruginosa</i> infection. This study aimed to investigate the prevalence and risk factors of of <i>P. aeruginosa</i> colonization in healthy humans. At the same time, the virulence of the isolated <i>P. aeruginosa</i> was also studied. In the study, 609 Vietnamese volunteers (310 females and 299 males, age range of 2 to 73 years), who had no acute infection or disease symptoms participated at the time of sample collection. Samples were taken from the throat, nostrils, and outer ears. <i>P. aeruginosa</i> was found in 19 participants (3.12%, 95% CI: 0.017–0.045), mainly from the throat (11/19, 57.89%). Participants with a history of sinusitis were 11.57 times more likely to be colonized with <i>P. aeruginosa</i> than participants without a history of sinusitis (OR: 11.57, 95% CI: 4.08–32.76, <i>p</i>-value < 0.0001, Fisher’s Exact test). Age and sex were not significantly associated with <i>P. aeruginosa</i> colonization. Among 16 <i>P. aeruginosa</i> isolates used in virulence tests, 100% (16/16) were positive for the synthesis of biofilm, pyocyanin, and siderophores; 93.75% (15/16) isolates were positive for the synthesis of gelatinase and protease; and 50% (8/16) isolates were positive for lipase. There were no differences in the pattern and range of virulence factors of <i>P. aeruginosa</i> isolates taken from participants with and without sinusitis history. <i>P. aeruginosa</i> colonized 3.12% of participants, and its presence was associated with sinusitis history.