Ureaplasma urealyticum and Mycoplasma hominis are associated with non-gonococcal urethritis, increased risk of recurrent miscarriage, infertility and pelvic inflammatory disease. Migration flows from other countries change the local epidemiological profile of infectious diseases of patients treated by general practitioners and hospital doctors. Few studies have evaluated this ever-changing issue in the Italian population. The aim of this study was to assess possible differences in prevalence and antimicrobial susceptibility of U. urealyticum and M. hominis in a population of 433 Italian and immigrant outpatients by means of the commercially available MYCOFAST(®) Screening EvolutioN 3 Kit. Prevalence of positive samples was 44.5% in Italian patients and 53.4% in immigrants. Samples positive for U. urealyticum and total isolates were more frequent in African patients: U. urealyticum, 51.5% vs 33.3%; Yates-corrected chi-square=3.98; p=0.046; total isolates, 54.5% vs 34.3%; Yates-corrected chi-square=4.45; p=0.035. Among samples positive for U. urealyticum, 66.4% were resistant to ciprofloxacin, whereas 27.6% to ofloxacin. In M. hominis isolates, 66.7% were resistant to both azythromycin and roxythromycin. Our study showed how prevalence of genital mycoplasmas and antibiotic resistance profiles change in relation to the country of origin. Therefore, surveillance is critical for the early cure and prevention of the occurrence of resistant strains.