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Updated: Aug 4, 2026
Optimized PCR-based Detection of Mycoplasma
Published on: June 20, 2011
A novel polymerase chain reaction assay to detect Mycoplasma genitalium
K Eastick1, J P Leeming, E O Caul
1Public Health Laboratory, Level 8, Bristol Royal Infirmary, Maudlin Street, Bristol BS2 8HW, UK. eastick@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.
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