Determination of antimicrobial sensitivity helps establish adequate treatment and avoids future genital tract diseases in women of fertile age. In Cuba, prevalence of mycoplasma in patients with vaginal discharge is unknown. The objective of this research was to determine frequency and antimicrobial sensitivity of Ureaplasma urealyticum and Mycoplasma hominis in women with vaginal discharge through analysis of laboratory data from vaginal smears from 255 patients referred to the Municipal Hygiene and Epidemiology Center in Güines, Mayabeque Province, Cuba. Mycoplasma System Plus (Italy) was used for detection, identification, count and sensitivity testing. The finding of mycoplasmas in almost two thirds of specimens examined suggests that the sexually active female population should be screened for these bacteria and that barrier contraception methods should be promoted to decrease their spread and prevent longterm sequelae. Such updating of local patterns of antimicrobial resistance supports decision making for best treatment options in patients with these infections. Our results should help clinicians in our area choose an antibiotic, and also confirm the utility of Mycoplasma System Plus for mycoplasma research in resource-scarce settings, to benefit individual and population health.