The aim of this paper is to describe the prevalence ofMycoplasma genitaliumandTrichomonas vaginalisin patients who visited general practitioners in the Netherlands. Additionally, we describe the prevalence ofM. genitaliumresistance to azithromycin and moxifloxacin. We used data from 7,411 consecutive female patients who were screened forChlamydia trachomatis,Neisseria gonorrhoeae,M. genitalium, andT. vaginalisand data from 5,732 consecutive male patients screened forC. trachomatis,N. gonorrhoeae, andM. genitalium. The prevalence ofM. genitaliumandT. vaginalisin female patients was 6.7% (95% CI: 6.2 to 7.4) and 1.9% (95%CI: 1.6 to 2.2%), respectively.M. genitaliumprevalence in male patients was 3.7% (3.3 to 4.3).M. genitaliumco-occurred withC. trachomatisin 1.4% (0.3 to 0.6%) of female and in 0.7% (0.5 to 0.9) of male patients. Macrolide resistance gene mutations and fluoroquinolone resistance gene mutations were detected in 73.8% and 9.9%, respectively. We concluded thatM.genitaliumis relatively infrequently found in a large general practitioner population in the Netherlands. It can co-occur withC. trachomatis, and is often resistant to azithromycin. Therefore, when treating sexually transmitted infections, these prevalence and resistance data should be taken into account.