Protocol for the detection of Mycoplasma genitalium by PCR from clinical specimens and subsequent detection of macrolide resistance-mediating mutations in region V of the 23S rRNA gene

Methods Mol Biol. 2012;903:129-39. doi: 10.1007/978-1-61779-937-2_8.

Abstract

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.

MeSH terms

  • Azithromycin / pharmacology
  • Drug Resistance, Bacterial / genetics*
  • Female
  • Humans
  • Macrolides / pharmacology*
  • Male
  • Molecular Diagnostic Techniques
  • Mutation*
  • Mycoplasma genitalium / drug effects
  • Mycoplasma genitalium / genetics*
  • Mycoplasma genitalium / isolation & purification*
  • Polymerase Chain Reaction / methods*
  • RNA, Bacterial / genetics
  • RNA, Ribosomal, 23S / genetics*
  • Taq Polymerase / metabolism

Substances

  • Macrolides
  • RNA, Bacterial
  • RNA, Ribosomal, 23S
  • Azithromycin
  • Taq Polymerase