Mycoplasma genitalium is an established cause of male nongonococcal urethritis, in particular in cases with recurrent disease and in those negative for Chlamydia trachomatis. In women M. genitalium causes cervicitis and there is increasing evidence that it is causing pelvic inflammatory disease (PID). Nucleic acid amplification tests are currently the only available methods for detection, but no commercially available tests have been thoroughly evaluated. Here we describe a TaqMan-based PCR test for detection of the infection and a conventional PCR that serves as a confirmatory assay with the possibility of sequencing the product for detection of macrolide resistance-mediating mutations.