This study examined the role of P fimbriae in the establishment of bacteriuria. Patients (n = 17) were subjected to intravesical inoculation with an asymptomatic bacteriuria strain, Escherichia coli 83972, or its P‐fimbriated (pap⁺/prs⁺) transformants. As shown by groupwise analysis, the pap⁺/prs⁺ transformants established bacteriuria more rapidly than E. coli 83972 (P = 0.021) and required a lower number of inoculations to reach 10⁵ cfu ml⁻¹ (P = 0.018). Intraindividual analysis showed that the pap⁺/prs⁺ transformants established bacteriuria more rapidly than E. coli 83972 in the patients who subsequently became carriers of both strains. Finally, bacterial establishment was shown to vary with the in vivo expression of P fimbriae. Bacterial counts were higher when P‐fimbrial expression was detected than when the pap⁺/prs⁺ strain showed a negative phenotype. The results suggested that P fimbriae enhance the establishment of bacteriuria and fulfil the molecular Koch postulates as a colonization factor in the human urinary tract.