Use of TaqMan 5' nuclease real-time PCR for quantitative detection of Mycoplasma genitalium DNA in males with and without urethritis who were attendees at a sexually transmitted disease clinic.
Mycoplasma genitalium is a cause of nongonococcal urethritis, particularly in patients not infected with Chlamydia trachomatis. A quantitative 5' nuclease assay (TaqMan PCR) was developed and validated. The assay detected a fragment of the MgPa adhesin gene by use of a TaqMan MGB (minor groove binder) probe and included an internal processing control to detect PCR inhibition. Urethral swab specimens and first-void urine samples from M. genitalium-positive men were examined, and the M. genitalium DNA load was correlated to symptoms and signs. The assay consistently detected <5 genome copies without cross-reactions with other mycoplasmas. Urine and urethral swab specimens from men with urethritis had higher M. genitalium DNA loads than specimens from men without urethritis. However, a very broad overlap of DNA loads between patients with and without urethritis was observed. Urethral swab specimens from patients with urethral discharge had a significantly higher DNA load than specimens from patients without discharge. This correlation was not found in first-void urine specimens.
Jensen
Jørgen Skov
JS
Mycoplasma Laboratory, Statens Serum Institut, DK-2300 Copenhagen S, Denmark. jsj@ssi.dk
Björnelius
Eva
E
Dohn
Birthe
B
Lidbrink
Peter
P
D001483:Base Sequence; D016001:Confidence Intervals; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification; D016680:Genome, Bacterial; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009175:Mycoplasma Infections / Q000175:diagnosis* / Q000453:epidemiology; D045704:Mycoplasma genitalium / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012749:Sexually Transmitted Diseases / Q000175:diagnosis / Q000453:epidemiology / Q000382:microbiology*; D013048:Specimen Handling; D013548:Sweden / Q000453:epidemiology; D014521:Urethra / Q000382:microbiology; D014526:Urethritis / Q000175:diagnosis / Q000209:etiology / Q000382:microbiology*
D017931:DNA Primers; D004269:DNA, Bacterial