ORIGINAL ARTICLE

A novel polymerase chain reaction assay to detect Mycoplasma genitalium

K Eastick¹, J P Leeming¹, E O Caul², P J Horner³ and M R Millar¹

¹ Public Health Laboratory, Level 8, Bristol Royal Infirmary, Maudlin Street, Bristol BS2 8HW, UK
² Public Health Laboratory, Myrtle Road, Kingsdown, Bristol BS2 8EL, UK
³ The Milne Centre for Sexual Health, Bristol Royal Infirmary, Maudlin Street, Bristol BS2 8HW, UK

Correspondence to:
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

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.

Keywords: Mycoplasma genitalium; polymerase chain reaction; urethritis

Abbreviations: HPF, high power field; IDEIA, amplified enzyme immunoassay; NGU, non-gonococcal urethritis; PCR, polymerase chain reaction; PMNL, polymorphonuclear leucocyte

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