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A novel polymerase chain reaction assay to detect Mycoplasma genitalium
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  1. K Eastick1,
  2. J P Leeming1,
  3. E O Caul2,
  4. P J Horner3,
  5. M R Millar1
  1. 1Public Health Laboratory, Level 8, Bristol Royal Infirmary, Maudlin Street, Bristol BS2 8HW, UK
  2. 2Public Health Laboratory, Myrtle Road, Kingsdown, Bristol BS2 8EL, UK
  3. 3The Milne Centre for Sexual Health, Bristol Royal Infirmary, Maudlin Street, Bristol BS2 8HW, UK
  1. Correspondence to:
 Dr K Eastick, Royal Infirmary of Edinburgh, Regional Clinical Virology Laboratory, Little France, Old Dalkeith Road, Edinburgh, EH16 4SU, UK
 eastick{at}btinternet.com

Abstract

Aims: To design and validate a polymerase chain reaction (PCR) assay targeting the 16S rRNA gene of Mycoplasma genitalium.

Methods: Primers were designed that were complementary to the 16S rRNA gene sequence of M genitalium. After optimisation of the reaction conditions, the PCR was tested against nine M genitalium strains, a dilution series of M genitalium DNA, and a panel of common microorganisms. The PCR was also challenged in parallel with a published assay against 54 urine specimens from men with urethritis.

Results: The expected 341 bp product was produced on amplification of material from all M genitalium strains and from none of the other microorganisms tested. The lower limit of detection was 50 genome copies. The new assay detected M genitalium DNA in nine of 54 men with urethritis, in comparison with eight positive specimens detected with the alternative PCR.

Conclusions: This novel PCR targeting the M genitalium 16S rRNA gene has been optimised and now provides a sensitive and specific alternative or addition to the available MgPa gene targeting assays.

  • Mycoplasma genitalium
  • polymerase chain reaction
  • urethritis
  • HPF, high power field
  • IDEIA, amplified enzyme immunoassay
  • NGU, non-gonococcal urethritis
  • PCR, polymerase chain reaction
  • PMNL, polymorphonuclear leucocyte
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