A novel polymerase chain reaction assay to detect Mycoplasma genitalium.
OBJECTIVE
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.
Eastick
K
K
Public Health Laboratory, Level 8, Bristol Royal Infirmary, Maudlin Street, Bristol BS2 8HW, UK. eastick@btinternet.com
Leeming
J P
JP
Caul
E O
EO
Horner
P J
PJ
Millar
M R
MR
D002692:Chlamydia trachomatis / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000032:analysis; D006801:Humans; D008297:Male; D009174:Mycoplasma / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D015203:Reproducibility of Results; D014526:Urethritis / Q000382:microbiology / Q000652:urine
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S